Diacylglycerol acyl transferase 2 inhibitors

ABSTRACT

Compounds of Formula I that inhibit the activity of the diacylglycerol acyltransferase 2 (DGAT2) and their uses in the treatment of diseases linked thereto in animals are described herein.

FIELD OF THE INVENTION

The present invention relates to new pharmaceutical compounds,pharmaceutical compositions containing these compounds, and their use toinhibit the activity of the diacylglycerol acyltransferase 2 (DGAT2).

BACKGROUND OF THE INVENTION

Triglycerides or triacylglycerols (TAG) represent a major form of energystorage in mammals. TAG's are formed by the sequential esterification ofglycerol with three fatty acids of varying chain lengths and degrees ofsaturation (1). TAG synthesized in the intestine or liver are packagedinto chylomicrons or very low-density lipoprotein (VLDL), respectively,and exported to peripheral tissues where they are hydrolysed to theirconstituent fatty acids and glycerol by lipoprotein lipase (LPL). Theresultant non-esterified fatty acids (NEFA) can either be metabolisedfurther to produce energy or reesterified and stored.

Under normal physiological conditions, the energy-dense TAG remainssequestered in various adipose depots until there is a demand for itsrelease, whereupon, it is hydrolyzed to glycerol and free fatty acidswhich are then released into the blood stream. This process is tightlyregulated by the opposing actions of insulin and hormones such ascatecholamines which promote the deposition and mobilization of TAGstores under various physiological conditions. In the post-prandialsetting, insulin acts to inhibit lipolysis, thereby, restraining therelease of energy in the form of NEFA and ensuring the appropriatestorage of dietary lipids in adipose depots. However, in patients withtype 2 diabetes, the ability of insulin to suppress lipolysis isameliorated and NEFA flux from adipocytes is inappropriately elevated.This, in turn, results in increased delivery of lipid to tissues such asmuscle and liver. In the absence of energetic demand the TAG and otherlipid metabolites, such as diacylglycerol (DAG) can accumulate and causea loss of insulin sensitivity (2). Insulin resistance in muscle ischaracterized by reduced glucose uptake and glycogen storage, whilst inthe liver, loss of insulin signaling gives rise to dysregulated glucoseoutput and over-production of TAG-rich VLDL, a hallmark of type 2diabetes (3). Elevated secretion of TAG-enriched VLDL, so called VLDL1particles, is thought to stimulate the production of small, denselow-density lipoprotein (sdLDL), a proatherogenic subfraction of LDLthat is associated with elevated risk of coronary heart disease (4).

Diacylglycerol acyltransferases (DGAT) catalyze the terminal step in TAGsynthesis, specifically, the esterification of a fatty acid withdiacylglycerol resulting in the formation of TAG. In mammals, two DGATenzymes (DGAT1 and DGAT2) have been characterized. Although theseenzymes catalyze the same enzymatic reaction their respective amino acidsequences are unrelated and they occupy distinct gene families. Miceharboring a disruption in the gene encoding DGAT1 are resistant todiet-induced obesity and have elevated energy expenditure and activity(5). Dgat1−/− mice exhibit dysregulated postaborpative release ofchylomicrons and accumulate lipid in the enterocytes (6). Themetabolically favorable phenotype observed in these mice is suggested tobe driven by loss of DGAT1 expression in the intestine (7). Importantly,despite a defect in lactation in female Dgat1−/− mice, these animalsretain the capacity to synthesize TAG suggesting the existence ofadditional DGAT enzymes. This observation and the isolation of a secondDGAT from the fungus Mortierella rammaniana led to the identificationand characterization of DGAT2 (8).

DGAT2 is highly expressed in liver and adipose, and unlike DGAT1,exhibits exquisite substrate specificity for DAG (8). Deletion of theDGAT2 gene in rodents results in defective intraunterine growth, severelipemia, impaired skin barrier function, and early post-natal death (9).Due to the lethality caused by loss of DGAT2, much of our understandingof the physiological role of DGAT2 derives from studies performed withantisense oligonucleotides (ASO) in rodent models of metabolic disease.In this setting, inhibition of hepatic DGAT2 resulted in improvements inplasma lipoprotein profile (decrease in total cholesterol and TAG) and areduction of hepatic lipid burden which was accompanied by improvedinsulin sensitivity and whole-body glucose control (10-12). Although themolecular mechanisms underlying these observations are not fullyelucidated, it is clear that suppression of DGAT2 results in adown-regulation of the expression of multiple genes encoding proteinsinvolved in lipogensis, including sterol regulatory element-bindingproteins 1c (SREBP1c) and stearoyl CoA-desaturase 1 (SCD1) (11, 12). Inparallel, oxidative pathways are induced as evidenced by increasedexpression of genes such as carnitine palmitoyl transfersase 1 (CPT1)(11). The net result of these changes is to decrease the levels ofhepatic DAG and TAG lipid which, in turn, leads to improved insulinresponsiveness in the liver. Furthermore, DGAT2 inhibition suppresseshepatic VLDL TAG secretion and reduction in circulating cholesterollevels. Finally, plasma apolipoprotein B (APOB) levels were suppressed,possibly due to decreased supply of TAG for lipidation of the newlysynthesized APOB protein (10, 12). The beneficial effects of DGAT2inhibition on both glycemic control and plasma cholesterol profilesuggest that this target might be valuable in the treatment of metabolicdisease (11). In addition, the observation that suppression of DGAT2activity results in reduced hepatic lipid accumulation suggests thatinhibitors of this enzyme might have utility in the treatment ofnon-alcoholic steatohepatitis (NASH), a highly prevalent liver diseasecharacterized by the deposition of excess fat in the liver.

In recent years, several small molecule inhibitors of DGAT2 have beenreported in literature (13-19) and patent applications (WO2013150416,WO2013137628, US20150259323, WO2015077299, WO2016036633, WO2016036638,WO2016036636).

-   1. Coleman, R. A., and D. G. Mashek. 2011. Chem Rev 111: 6359-6386.-   2. Erion, D. M., and G. I. Shulman. 2010. Nat Med 16: 400-402.-   3. Choi, S. H., and H. N. Ginsberg. 2011. Trends Endocrinol Metab    22: 353-363.-   4. St-Pierre, A. C. et. al. 2005. Arterioscler Thromb Vasc Biol 25:    553-559.-   5. Smith, S. J. et. al. 2000. Nat Genet 25: 87-90.-   6. Buhman, K. K. et. al. 2002. J Biol Chem 277: 25474-25479.-   7. Lee, B., A. M. et. al. 2010. J Lipid Res 51: 1770-1780.-   8. Yen, C. L. et. al. 2008. J Lipid Res 49: 2283-2301.-   9. Stone, S. J. et. al. 2004. J Biol Chem 279: 11767-11776.-   10. Liu, Y. et. al. 2008. Biochim Biophys Acta 1781: 97-104.-   11. Choi, C. S. et. al. 2007. J Biol Chem 282: 22678-22688.-   12. Yu, X. X. et. al. 2005. Hepatology 42: 362-371.-   13. Qi, J. et. al. J. Lipid. Res. 2012, 53 (6), 1106-16.-   14. Wurie, H. R. et. al. FEBS. J. 2012, 279 (17), 3033-47;-   15. Kim, M. O. et. al. Biol. Pharm. Bull. 2013, 36 (7), 1167-73-   16. Lee, K. et. al. Org. Biomol. Chem. 2013, 11 (5), 849-58-   17. Kim, M. O. et. al. Biol. Pharm. Bull. 2014, 37 (10), 1655-1660.-   18. Futatsugi, K. et. al. J Med Chem 2015, 58 (18), 7173-85.-   19. Imbriglio, J. E. et. al. J. Med. Chem. 2015, 58 (23), 9345-9353.

SUMMARY OF THE INVENTION

The present application is directed at compounds of Formula (I) and (Ia)

wherein

D¹ and D² are each independently N or CH;

R¹ is H, or (C₁-C₂)alkyl optionally substituted with one or twosubstituents each independently selected from fluoro and(C₃-C₆)cycloalkyl;

R² is H or fluoro;

R³ is

R⁴ is H, cyano, or (C₁-C₄)alkyl optionally substituted with one or twosubstituents each independently selected from —OH and —S(O)₂R⁶;

R⁵ is H or —OH; and

R⁶ is (C₁-C₄)alkyl;

or a pharmaceutically acceptable salt thereof.

The present invention is also directed at a crystal comprising acompound having the structure:

or a pharmaceutically acceptable salt thereof.

The present invention is also directed at pharmaceutical compositionsthat include a compound of Formula (I) or (Ia) or a pharmaceuticallyacceptable salt of said compound, present in a therapeutically effectiveamount, in admixture with at least one pharmaceutically acceptableexcipient.

Furthermore, the present invention is directed at pharmaceuticalcompositions that include a compound of Formula (I) or (Ia) or apharmaceutically acceptable salt of said compound, present in atherapeutically effective amount, in admixture with at least onepharmaceutically acceptable excipient and further including at least oneadditional pharmaceutical agent selected from the group consisting of ananti-inflammatory agent, an anti-diabetic agent, and a cholesterol/lipidmodulating agent.

In another embodiment, the method of the present invention is for thetreatment of hyperlipidemia, Type I diabetes, Type II diabetes mellitus,idiopathic Type I diabetes (Type Ib), latent autoimmune diabetes inadults (LADA), early-onset Type 2 diabetes (EOD), youth-onset atypicaldiabetes (YOAD), maturity onset diabetes of the young (MODY),malnutrition-related diabetes, gestational diabetes, coronary heartdisease, ischemic stroke, restenosis after angioplasty, peripheralvascular disease, intermittent claudication, myocardial infarction,dyslipidemia, post-prandial lipemia, conditions of impaired glucosetolerance (IGT), conditions of impaired fasting plasma glucose,metabolic acidosis, ketosis, arthritis, obesity, osteoporosis,hypertension, congestive heart failure, left ventricular hypertrophy,peripheral arterial disease, diabetic retinopathy, macular degeneration,cataract, diabetic nephropathy, glomerulosclerosis, chronic renalfailure, diabetic neuropathy, metabolic syndrome, syndrome X,premenstrual syndrome, angina pectoris, thrombosis, atherosclerosis,transient ischemic attacks, stroke, vascular restenosis, hyperglycemia,hyperinsulinemia, hypertrygliceridemia, insulin resistance, impairedglucose metabolism, erectile dysfunction, skin and connective tissuedisorders, foot ulcerations and ulcerative colitis, endothelialdysfunction and impaired vascular compliance, hyper apo Blipoproteinemia, Alzheimer's, schizophrenia, impaired cognition,inflammatory bowel disease, ulcerative colitis, Crohn's disease, andirritable bowel syndrome, non-alcoholic steatohepatitis (NASH), ornon-alcoholic fatty liver disease (NAFLD), in humans.

In another embodiment, the method reduces portal hypertension, hepaticprotein synthetic capability, hyperbilirubinemia, or encephalopathy.

The present invention is also directed at a method for the treatment ofreduction of at least one point in severity of nonalcoholic fatty liverdisease or nonalcoholic steatohepatitis grading scoring systems,reduction of the level of serum markers of nonalcoholic steatohepatitisactivity, reduction of nonalcoholic steatohepatitis disease activity orreduction in the medical consequences of nonalcoholic steatohepatitis inhumans comprising the step of administering to a human in need of suchreduction diabetes comprising the administration of an effective aneffective amount of a compound of Formula (I) or (Ia) or apharmaceutically acceptable salt of said compound or a pharmaceuticallyacceptable salt of said compound to a patient in need thereof.

The present invention is also directed at a method for treating fattyliver, nonalcoholic fatty liver disease, nonalcoholic steatohepatitis,nonalcoholic steatohepatitis with liver fibrosis, nonalcoholicsteatohepotitis with cirrhosis, or nonalcoholic steatohepatitis withcirrhosis and hepatocellular carcinoma metabolic or metabolic-relateddisease, condition or disorder in humans comprising the step ofadministering to a human in need of such treatment comprising the stepof administering to a patient a therapeutically effective amount of acompound of Formula (I) or (Ia) or a pharmaceutically acceptable salt ofsaid compound or a pharmaceutically acceptable salt of said compound.

The present invention is also directed at a method for treating fattyliver, nonalcoholic fatty liver disease, nonalcoholic steatohepatitis,nonalcoholic steatohepatitis with liver fibrosis, nonalcoholicsteatohepotitis with cirrhosis, or nonalcoholic steatohepatitis withcirrhosis and hepatocellular carcinoma metabolic or metabolic-relateddisease, condition or disorder in humans comprising the step ofadministering to a human in need of such treatment comprising the stepof administering to a patient in need of such treatment atherapeutically effective amount of two separate pharmaceuticalcompositions comprising

(i) a first composition that includes a compound of Formula (I) or (Ia)or a pharmaceutically acceptable salt of said compound, present in atherapeutically effective amount, in admixture with at least onepharmaceutically acceptable excipient; and(ii) a second composition comprising at least one additionalpharmaceutical agent selected from the group consisting of ananti-inflammatory agent, an anti-diabetic agent, and a cholesterol/lipidmodulating agent and an anti-diabetic agent, and at least onepharmaceutically acceptable excipient.

It is to be understood that both the foregoing general description andthe following detailed description are exemplary and explanatory onlyand are not restrictive of the invention, as claimed.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a characteristic x-ray powder diffraction pattern showingcrystalline Form 1 of Example 1 (Vertical Axis: Intensity (CPS);Horizontal Axis: Two theta (degrees)).

FIG. 2 is a characteristic x-ray powder diffraction pattern showingcrystalline Form 2 of Example 1 (Vertical Axis: Intensity (CPS);Horizontal Axis: Two theta (degrees)).

FIGS. 3 and 4 summarize the effects of oral administration with Example1 on plasma and hepatic triglyceride levels in western diet fed SpragueDawley rats respectively.

DETAILED DESCRIPTION OF THE INVENTION

The present invention may be understood more readily by reference to thefollowing detailed description of exemplary embodiments of the inventionand the examples included therein.

It is to be understood that this invention is not limited to specificsynthetic methods of making that may of course vary. It is also to beunderstood that the terminology used herein is for the purpose ofdescribing particular embodiments only and is not intended to belimiting. In this specification and in the claims that follow, referencewill be made to a number of terms that shall be defined to have thefollowing meanings:

As used herein in the specification, “a” or “an” may mean one or more.As used herein in the claim(s), when used in conjunction with the word“comprising”, the words “a” or “an” may mean one or more than one. Asused herein “another” may mean at least a second or more.

The term “about” refers to a relative term denoting an approximation ofplus or minus 10% of the nominal value it refers, in one embodiment, toplus or minus 5%, in another embodiment, to plus or minus 2%. For thefield of this disclosure, this level of approximation is appropriateunless the value is specifically stated to require a tighter range.

“Compounds” when used herein includes any pharmaceutically acceptablederivative or variation, including conformational isomers (e.g., cis andtrans isomers) and all optical isomers (e.g., enantiomers anddiastereomers), racemic, diastereomeric and other mixtures of suchisomers, as well as solvates, hydrates, isomorphs, polymorphs,tautomers, esters, salt forms, and prodrugs. The expression “prodrug”refers to compounds that are drug precursors which followingadministration, release the drug in vivo via some chemical orphysiological process (e.g., a prodrug on being brought to thephysiological pH or through enzyme action is converted to the desireddrug form). Exemplary prodrugs upon cleavage release the correspondingfree acid, and such hydrolyzable ester-forming residues of the compoundsof the present invention include but are not limited to those having acarboxyl moiety wherein the free hydrogen is replaced by (C₁-C₄)alkyl,(C₂-C₇)alkanoyloxymethyl, 1-(alkanoyloxy)ethyl having from 4 to 9 carbonatoms, 1-methyl-1-(alkanoyloxy)-ethyl having from 5 to 10 carbon atoms,alkoxycarbonyloxymethyl having from 3 to 6 carbon atoms,1-(alkoxycarbonyloxy)ethyl having from 4 to 7 carbon atoms,1-methyl-1-(alkoxycarbonyloxy)ethyl having from 5 to 8 carbon atoms,N-(alkoxycarbonyl)aminomethyl having from 3 to 9 carbon atoms,1-(N-(alkoxycarbonyl)amino)ethyl having from 4 to 10 carbon atoms,3-phthalidyl, 4-crotonolactonyl, gamma-butyrolacton-4-yl,di-N,N—(C₁-C₂)alkylamino(C₂-C₃)alkyl (such as β-dimethylaminoethyl),carbamoyl-(C₁-C₂)alkyl, N, N-di(C₁-C₂)alkylcarbamoyl-(C₁-C₂)alkyl andpiperidino-, pyrrolidino- or morpholino(C₂-C₃)alkyl.

As used herein, an arrowhead, “

” or wavy line, “

” denotes a point of attachment of a substituent to another group.

By “alkyl” is meant straight chain saturated hydrocarbon or branchedchain saturated hydrocarbon. Exemplary of such alkyl groups (assumingthe designated length encompasses the particular example) are methyl,ethyl, propyl, isopropyl, butyl, sec-butyl, tertiary butyl, isobutyl,pentyl, isopentyl, neopentyl, tertiary pentyl, 1-methylbutyl,2-methylbutyl, 3-methylbutyl, hexyl, isohexyl, heptyl and octyl.

The term “aryl” means a carbocyclic aromatic system containing one, twoor three rings wherein such rings may be fused. If the rings are fused,one of the rings must be fully unsaturated and the fused ring(s) may befully saturated, partially unsaturated or fully unsaturated. The term“fused” means that a second ring is present (ie, attached or formed) byhaving two adjacent atoms in common (ie, shared) with the first ring.The term “fused” is equivalent to the term “condensed”. The term “aryl”embraces aromatic radicals such as phenyl, naphthyl, tetrahydronaphthyl,indanyl, biphenyl, benzo[b][1,4]oxazin-3(4H)-onyl, 2,3-dihydro-1Hindenyl, and 1,2,3,4-tetrahydronaphthalenyl.

“Cycloalkyl” refers to a nonaromatic ring that is fully hydrogenatedhaving one, two or three rings wherein such rings may be fused, whereinfused is defined above. Cycloalkyl also includes bicyclic structuresthat may be bridged or spirocyclic in nature with each individual ringwithin the bicycle varying from 3-8 atoms. Examples of such carbocyclicrings include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

The term “heteroaryl” means an aromatic carbocyclic system containingone, two, three or four heteroatoms selected independently from oxygen,nitrogen and sulfur and having one, two or three rings wherein suchrings may be fused, wherein fused is defined above. The term“heteroaryl” includes but is not limited to furyl, thienyl, oxazolyl,thiazolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, isoxazolyl,isothiazolyl, oxadiazolyl, thiadiazolyl, pyridinyl, pyridiazinyl,pyrimidinyl, pyrazinyl, pyridin-2(1H)-onyl, pyridazin-2(1H)-onyl,pyrimidin-2(1H)-onyl, pyrazin-2(1H)-onyl, imidazo[1,2-a]pyridinyl,pyrazolo[1,5-a]pyridinyl, 5,6,7,8-tetrahydroisoquinolinyl,5,6,7,8-tetrahydroquinolinyl, 6,7-dihydro-5H-cyclopenta[b]pyridinyl,6,7-dihydro-5H-cyclopenta[c]pyridinyl,1,4,5,6-tetrahydrocyclopenta[c]pyrazolyl,2,4,5,6-tetrahydrocyclopenta[c]pyrazolyl,5,6-dihydro-4H-pyrrolo[1,2-b]pyrazolyl,6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazolyl,5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyridinyl,4,5,6,7-tetrahydropyrazolo[1,5-a]pyridinyl,4,5,6,7-tetrahydro-1H-indazolyl and 4,5,6,7-tetrahydro-2H-indazolyl.

It is to be understood that if a carbocyclic or heterocyclic moiety maybe bonded or otherwise attached to a designated substrate throughdiffering ring atoms without denoting a specific point of attachment,then all possible points are intended, whether through a carbon atom or,for example, a trivalent nitrogen atom. For example, the term “pyridyl”means 2-, 3- or 4-pyridyl, the term “thienyl” means 2- or 3-thienyl, andso forth.

“Patient” refers to warm blooded animals such as, for example, guineapigs, mice, rats, gerbils, cats, rabbits, dogs, cattle, goats, sheep,horses, monkeys, chimpanzees, and humans.

By “pharmaceutically acceptable” is meant that the substance orcomposition must be compatible chemically and/or toxicologically, withthe other ingredients comprising a formulation, and/or the mammal beingtreated therewith.

As used herein, the expressions “reaction-inert solvent” and “inertsolvent” refer to a solvent or a mixture thereof which does not interactwith starting materials, reagents, intermediates or products in a mannerwhich adversely affects the yield of the desired product.

As used herein, the term “selectivity” or “selective” refers to agreater effect of a compound in a first assay, compared to the effect ofthe same compound in a second assay. For example, in “gut selective”compounds, the first assay is for the half life of the compound in theintestine and the second assay is for the half life of the compound inthe liver.

“Therapeutically effective amount” means an amount of a compound of thepresent invention that (i) treats or prevents the particular disease,condition, or disorder, (ii) attenuates, ameliorates, or eliminates oneor more symptoms of the particular disease, condition, or disorder, or(iii) prevents or delays the onset of one or more symptoms of theparticular disease, condition, or disorder described herein.

The term “treating”, “treat” or “treatment” as used herein embraces bothpreventative, i.e., prophylactic, and palliative treatment, i.e.,relieve, alleviate, or slow the progression of the patient's disease (orcondition) or any tissue damage associated with the disease.

The compounds of the present invention may contain asymmetric or chiralcenters, and, therefore, exist in different stereoisomeric forms. Unlessspecified otherwise, it is intended that all stereoisomeric forms of thecompounds of the present invention as well as mixtures thereof,including racemic mixtures, form part of the present invention. Inaddition, the present invention embraces all geometric and positionalisomers. For example, if a compound of the present inventionincorporates a double bond or a fused ring, both the cis- andtrans-forms, as well as mixtures, are embraced within the scope of theinvention.

Chiral compounds of the invention (and chiral precursors thereof) may beobtained in enantiomerically-enriched form using chromatography,typically high pressure liquid chromatography (HPLC) or supercriticalfluid chromatography (SFC), on a resin with an asymmetric stationaryphase and with a mobile phase consisting of a hydrocarbon, typicallyheptane or hexane, containing from 0 to 50% isopropanol, typically from2 to 20%, and from 0 to 5% of an alkylamine, typically 0.1% diethylamine(DEA) or isopropylamine. Concentration of the eluent affords theenriched mixture.

Diastereomeric mixtures can be separated into their individualdiastereoisomers on the basis of their physical chemical differences bymethods well known to those skilled in the art, such as bychromatography and/or fractional crystallization. Enantiomers can beseparated by converting the enantiomeric mixture into a diastereomericmixture by reaction with an appropriate optically active compound (e.g.chiral auxiliary such as a chiral alcohol or Mosher's acid chloride),separating the diastereoisomers and converting (e.g. hydrolyzing) theindividual diastereoisomers to the corresponding pure enantiomers.Enantiomers can also be separated by use of a chiral HPLC column.Alternatively, the specific stereoisomers may be synthesized by using anoptically active starting material, by asymmetric synthesis usingoptically active reagents, substrates, catalysts or solvents, or byconverting one stereoisomer into the other by asymmetric transformation.

Where the compounds of the present invention possess two or morestereogenic centers and the absolute or relative stereochemistry isgiven in the name, the designations R and S refer respectively to eachstereogenic center in ascending numerical order (1, 2, 3, etc.)according to the conventional IUPAC number schemes for each molecule.Where the compounds of the present invention possess one or morestereogenic centers and no stereochemistry is given in the name orstructure, it is understood that the name or structure is intended toencompass all forms of the compound, including the racemic form.

The compounds of this invention may contain olefin-like double bonds.When such bonds are present, the compounds of the invention exist as cisand trans configurations and as mixtures thereof. The term “cis” refersto the orientation of two substituents with reference to each other andthe plane of the ring (either both “up” or both “down”). Analogously,the term “trans” refers to the orientation of two substituents withreference to each other and the plane of the ring (the substituentsbeing on opposite sides of the ring).

It is also possible that the intermediates and compounds of the presentinvention may exist in different tautomeric forms, and all such formsare embraced within the scope of the invention. The term “tautomer” or“tautomeric form” refers to structural isomers of different energieswhich are interconvertible via a low energy barrier. For example, protontautomers (also known as prototropic tautomers) include interconversionsvia migration of a proton, such as keto-enol and imine-enamineisomerizations.

Valence tautomers include interconversions by reorganization of some ofthe bonding electrons.

Included within the scope of the claimed compounds present invention areall stereoisomers, geometric isomers and tautomeric forms of thecompounds of Formula (I), including compounds exhibiting more than onetype of isomerism, and mixtures of one or more thereof. Also includedare acid addition or base salts wherein the counterion is opticallyactive, for example, D-lactate or L-lysine, or racemic, for example,DL-tartrate or DL-arginine.

The present invention includes all pharmaceutically acceptableisotopically-labelled compounds of Formula (I) wherein one or more atomsare replaced by atoms having the same atomic number, but an atomic massor mass number different from the atomic mass or mass number usuallyfound in nature.

Examples of isotopes suitable for inclusion in the compounds of theinvention include isotopes of hydrogen, such as ²H and ³H, carbon, suchas ¹¹C, ¹³C and ¹⁴C, chlorine, such as ³⁶Cl, fluorine, such as ¹⁸F,iodine, such as ¹²³I, ¹²⁴I and ¹²⁵I, nitrogen, such as ¹³N and ¹⁵N,oxygen, such as ¹⁵O, ¹⁷O and ¹⁸O, phosphorus, such as ³²P, and sulphur,such as ³⁵S.

Certain isotopically-labelled compounds of Formula (I), for example,those incorporating a radioactive isotope, are useful in drug and/orsubstrate tissue distribution studies. The radioactive isotopes tritium,i.e. ³H, and carbon-14, i.e. ¹⁴C, are particularly useful for thispurpose in view of their ease of incorporation and ready means ofdetection.

Substitution with heavier isotopes such as deuterium, i.e. ²H, mayafford certain therapeutic advantages resulting from greater metabolicstability, for example, increased in vivo half-life or reduced dosagerequirements, and hence may be preferred in some circumstances.

Substitution with positron emitting isotopes, such as ¹¹C, ¹⁸F, ¹⁵O and¹³N, can be useful in Positron Emission Tomography (PET) studies forexamining substrate receptor occupancy.

Isotopically-labelled compounds of Formula (I) can generally be preparedby conventional techniques known to those skilled in the art or byprocesses analogous to those described in the accompanying Examples andPreparations using an appropriate isotopically-labelled reagents inplace of the non-labelled reagent previously employed.

The compounds of the present invention may be isolated and used per se,or when possible, in the form of its pharmaceutically acceptable salt.The term “salts” refers to inorganic and organic salts of a compound ofthe present invention. These salts can be prepared in situ during thefinal isolation and purification of a compound, or by separatelytreating the compound with a suitable organic or inorganic acid or baseand isolating the salt thus formed. The acids which are used to preparethe pharmaceutically acceptable acid addition salts of theaforementioned base compounds of this invention are those which formnon-toxic acid addition salts, (i.e., salts containing pharmacologicallyacceptable anions, such as the hydrochloride, hydrobromide, hydroiodide,nitrate, sulfate, bisulfate, phosphate, acid phosphate, acetate,lactate, citrate, acid citrate, tartrate, bitartrate, succinate,maleate, fumarate, gluconate, saccharate, benzoate, methanesulfonate,ethanesulfonate, benzenesulfonate, naphthylate, mesylate,glucoheptonate, lactobionate, laurylsulphonate, hexafluorophosphate,benzene sulfonate, tosylate, formate, trifluoroacetate, oxalate,besylate, palmitiate, pamoate, malonate, stearate, laurate, malate,borate, p-toluenesulfonate and pamoate (i.e.,1,1′-methylene-bis-(2-hydroxy-3-naphthoate)) salts.

The invention also relates to base addition salts of the compounds ofthe present invention. The chemical bases that may be used as reagentsto prepare pharmaceutically acceptable base salts of those compounds ofthe present invention that are acidic in nature are those that formnon-toxic base salts with such compounds. Such non-toxic base saltsinclude, but are not limited to those derived from suchpharmacologically acceptable cations such as alkali metal cations (e.g.,lithium, potassium and sodium) and alkaline earth metal cations (e.g.,calcium and magnesium), ammonium or water-soluble amine addition saltssuch as N-methylglucamine-(meglumine), tetramethylammonium,tetraethylammonium, methylamine, dimethylamine, trimethylamine,triethylamine, ethylamine, and the lower alkanolammonium and other basesalts of pharmaceutically acceptable organic amines. See e.g. Berge, etal. J. Pharm. Sci. 66, 1-19 (1977).

Certain compounds of the present invention may exist in more than onecrystal form (generally referred to as “polymorphs”). Polymorphs may beprepared by crystallization under various conditions, for example, usingdifferent solvents or different solvent mixtures for recrystallization;crystallization at different temperatures; and/or various modes ofcooling, ranging from very fast to very slow cooling duringcrystallization. Polymorphs may also be obtained by heating or meltingthe compound of the present invention followed by gradual or fastcooling. The presence of polymorphs may be determined by solid probe NMRspectroscopy, IR spectroscopy, differential scanning calorimetry, powderX-ray diffraction or such other techniques.

In one embodiment, R³ is

In another embodiment, R³ is

In a further embodiment, R¹ is methyl.

In yet another embodiment, R⁴ is H, —CH₂OH, or cyano.

In another embodiment, the compound is

-   (S)-2-(5-((3-Ethoxy-5-fluoropyridin-2-yl)oxy)pyridin-3-yl)-N-(tetrahydrofuran-3-yl)pyrimidine-5-carboxamide;-   N-(2-cyanopropan-2-yl)-2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)pyrimidine-5-carboxamide;-   2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(3-methyl-1,1-dioxidotetrahydrothiophen-3-yl)pyrimidine-5-carboxamide;-   2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(1-hydroxy-2-methylpropan-2-yl)pyrimidine-5-carboxamide;-   (S)-2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(tetrahydrofuran-3-yl)pyrimidine-5-carboxamide;-   (S)-2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(3-(hydroxymethyl)tetrahydrofuran-3-yl)pyrimidine-5-carboxamide;-   (R)-2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(3-(hydroxymethyl)tetrahydrofuran-3-yl)pyrimidine-5-carboxamide;-   2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(2-methyl-1-(methylsulfonyl)propan-2-yl)pyrimidine-5-carboxamide;-   (S)-2-(5-((3-(2-fluoroethoxy)pyridin-2-yl)oxy)pyridin-3-yl)-N-(tetrahydrofuran-3-yl)pyrimidine-5-carboxamide;-   3-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(1-hydroxy-2-methylpropan-2-yl)-1,2,4-triazine-6-carboxamide;-   N-(1,3-dihydroxy-2-methylpropan-2-yl)-2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)pyrimidine-5-carboxamide;-   (S)-3-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(tetrahydrofuran-3-yl)-1,2,4-triazine-6-carboxamide;-   N-(1,1-dioxidotetrahydrothiophen-3-yl)-2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)pyrimidine-5-carboxamide;-   (R)-2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(tetrahydrofuran-3-yl)pyrimidine-5-carboxamide;    or-   (S)2-(5-((3-ethoxypyrazin-2-yl)oxy)pyridin-3-yl)-N-(1-hydroxy-2-methylpropan-2-yl)pyrimidine-5-carboxamide;

or a pharmaceutically acceptable salt thereof.

In another embodiment, the compound is:

-   (R)-2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(3-(hydroxymethyl)tetrahydrofuran-3-yl)pyrimidine-5-carboxamide;-   (S)-2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(tetrahydrofuran-3-yl)pyrimidine-5-carboxamide;    or-   (S)-2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(3-(hydroxymethyl)tetrahydrofuran-3-yl)pyrimidine-5-carboxamide,    or a pharmaceutically acceptable salt thereof.

In a further embodiment, the compound has the structure:

or a pharmaceutically acceptable salt thereof.

In a further embodiment, the compound of Formula (I) or (Ia) or a saltof the compound is present in a pharmaceutical composition in atherapeutically effective amount, in admixture with at least onepharmaceutically acceptable excipient.

In a further embodiment, the composition further includes at least oneadditional pharmaceutical agent selected from the group consisting of ananti-inflammation agent, an anti-diabetic agent, and a cholesterol/lipidmodulating agent.

In an embodiment, the method for the treatment of diabetes includes theadministration of an effective amount of compound of the presentinvention or a pharmaceutically acceptable salt of said compound to apatient in need thereof.

In another embodiment, the method for treating a metabolic ormetabolic-related disease, condition or disorder includes the step ofadministering to a patient a therapeutically effective amount of acompound of the present invention or a pharmaceutically acceptable saltof said compound.

In another embodiment, the method for treating a condition selected fromthe group consisting of hyperlipidemia, Type I diabetes, Type IIdiabetes mellitus, idiopathic Type I diabetes (Type Ib), latentautoimmune diabetes in adults (LADA), early-onset Type 2 diabetes (EOD),youth-onset atypical diabetes (YOAD), maturity onset diabetes of theyoung (MODY), malnutrition-related diabetes, gestational diabetes,coronary heart disease, ischemic stroke, restenosis after angioplasty,peripheral vascular disease, intermittent claudication, myocardialinfarction (e.g. necrosis and apoptosis), dyslipidemia, post-prandiallipemia, conditions of impaired glucose tolerance (IGT), conditions ofimpaired fasting plasma glucose, metabolic acidosis, ketosis, arthritis,obesity, osteoporosis, hypertension, congestive heart failure, leftventricular hypertrophy, peripheral arterial disease, diabeticretinopathy, macular degeneration, cataract, diabetic nephropathy,glomerulosclerosis, chronic renal failure, diabetic neuropathy,metabolic syndrome, syndrome X, premenstrual syndrome, coronary heartdisease, angina pectoris, thrombosis, atherosclerosis, myocardialinfarction, transient ischemic attacks, stroke, vascular restenosis,hyperglycemia, hyperinsulinemia, hyperlipidemia, hypertrygliceridemia,insulin resistance, impaired glucose metabolism, conditions of impairedglucose tolerance, conditions of impaired fasting plasma glucose,obesity, erectile dysfunction, skin and connective tissue disorders,foot ulcerations and ulcerative colitis, endothelial dysfunction andimpaired vascular compliance, hyper apo B lipoproteinemia, Alzheimer's,schizophrenia, impaired cognition, inflammatory bowel disease,ulcerative colitis, Crohn's disease, and irritable bowel syndrome,non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease(NAFLD), includes the administration of an effective amount of acompound according to the present invention or a pharmaceuticallyacceptable salt of said compound.

In a further embodiment, the method for treating a metabolic ormetabolic-related disease, condition or disorder includes the step ofadministering to a patient in need of such treatment two separatepharmaceutical compositions comprising

(i) a first composition according to the present invention; and

(ii) a second composition comprising at least one additionalpharmaceutical agent selected from the group consisting of ananti-obesity agent and an anti-diabetic agent, and at least onepharmaceutically acceptable excipient.

In yet a further embodiment, the method of the present invention isperformed when said first composition and said second composition areadministered simultaneously.

In yet another embodiment, the method of the present invention isperformed when first composition and said second composition areadministered sequentially and in any order.

In one embodiment, when two compositions are administered, the firstcomposition and the second composition are administered simultaneously.In another embodiment, the first composition and the second compositionare administered sequentially and in any order.

Compounds of the present invention may be synthesized by syntheticroutes that include processes analogous to those well-known in thechemical arts, particularly in light of the description containedherein. The starting materials are generally available from commercialsources such as Aldrich Chemicals (Milwaukee, Wis.) or are readilyprepared using methods well known to those skilled in the art (e.g.,prepared by methods generally described in Louis F. Fieser and MaryFieser, Reagents for Organic Synthesis, v. 1-19, Wiley, New York(1967-1999 ed.), or Beilsteins Handbuch der organischen Chemie, 4, Aufl.ed. Springer-Verlag, Berlin, including supplements (also available viathe Beilstein online database)). Many of the compounds used herein, arerelated to, or are derived from compounds in which there is a largescientific interest and commercial need, and accordingly many suchcompounds are commercially available or are reported in the literatureor are easily prepared from other commonly available substances bymethods which are reported in the literature.

For illustrative purposes, the reaction schemes depicted below providepotential routes for synthesizing the compounds of the present inventionas well as key intermediates. For a more detailed description of theindividual reaction steps, see the Examples section below. Those skilledin the art will appreciate that other synthetic routes may be used tosynthesize the inventive compounds. Although specific starting materialsand reagents are discussed below, other starting materials and reagentscan be easily substituted to provide a variety of derivatives and/orreaction conditions. In addition, many of the compounds prepared by themethods described below can be further modified in light of thisdisclosure using conventional chemistry well known to those skilled inthe art.

In the preparation of the Formula I compounds it is noted that some ofthe preparation methods useful for the preparation of the compoundsdescribed herein may require protection of remote functionality (e.g.,primary amine, secondary amine, carboxyl in Formula I precursors). Theneed for such protection will vary depending on the nature of the remotefunctionality and the conditions of the preparation methods. The needfor such protection is readily determined by one skilled in the art. Theuse of such protection/deprotection methods is also within the skill inthe art. For a general description of protecting groups and their use,see T.W. Greene, Protective Groups in Organic Synthesis, John Wiley &Sons, New York, 1991.

For example, certain compounds contain primary amines or carboxylic acidfunctionalities which may interfere with reactions at other sites of themolecule if left unprotected. Accordingly, such functionalities may beprotected by an appropriate protecting group which may be removed in asubsequent step. Suitable protecting groups for amine and carboxylicacid protection include those protecting groups commonly used in peptidesynthesis (such as N-t-butoxycarbonyl (Boc), benzyloxycarbonyl (Cbz),and 9-fluorenylmethylenoxycarbonyl (Fmoc) for amines and lower alkyl orbenzyl esters for carboxylic acids) which are generally not chemicallyreactive under the reaction conditions described and can typically beremoved without chemically altering other functionality in the Formula Iand Ia compounds.

The Reaction Schemes described below are intended to provide a generaldescription of the methodology employed in the preparation of thecompounds of the present invention. Some of the compounds of the presentinvention contain a single chiral center. In the following Schemes, thegeneral methods for the preparation of the compounds are shown either inracemic or enantioenriched form. It will be apparent to one skilled inthe art that all of the synthetic transformations can be conducted in aprecisely similar manner whether the materials are enantioenriched orracemic. Moreover the resolution to the desired optically activematerial may take place at any desired point in the sequence using wellknown methods such as described herein and in the chemistry literature.

In the Reaction Schemes I and II, the variables D¹, D², R¹, R², and R³are as described in the summary except where otherwise noted. Variable Ris methyl or ethyl. Reaction Scheme I outlines general procedures thatcan be used to provide compounds of the present invention having Formula(I).

Compounds of Formula (I) may be synthesized starting from appropriateintermediates through methods described in the literature such as: J.Med. Chem., 2007, 50, 2990-3003; Monatsh Chem, 2012, 143, 1575-1592; J.Med. Chem., 2011, 54, 6342-6363; Org. Proc. Res. Dev. 2014, 18,1145-1152; Angew. Chem. Int. Ed. 2011, 50, 9943; J. Am. Chem. Soc. 2005,127, 8146; J. Org. Chem. 2008, 73, 284; Org. Lett. 2002, 4, 973; Org.Lett., 2011, 13, 1840-1843; Metal Catalyzed Cross-Coupling Reactions andMore, Wiley-VCH, Weinheim, Germany, 2014, 3, 995; Applications ofTransition Metal Catalysis in Drug Discovery and Development, John Wiley& Sons, inc., Hoboken, N.J., USA, 2012, 3, 97. Intermediates (1a) and(1b) are commercially available and/or may be prepared via methods knownto those skilled in the art. For example, intermediates (1a) and (1b)may be synthesized through methods described in the literature such as:J. Med. Chem. 2000, 43, 3995; Org. Proc. Res. Dev. 2010, 14, 936.Intermediates (2a) and (2b) are commercially available or are describedin the literature and may be prepared via methods known to those skilledin the art, including those described below (Reaction Scheme II).

Intermediate (3a) may be prepared from intermediates (1a) and (2a) in atransition metal mediated coupling reaction. One of the halides (1a) or(2a) may be converted to an organometallic reagent, such as a boronicacid, zincate, stannane, or Grignard derivative using methods well knownto those skilled in the art. The resulting organometallic reagent maythen be reacted with the other halide intermediate in a transition metalcatalyzed cross coupling reaction. Preferably, intermediate (2a) isconverted to a zincate and is coupled to intermediate (1a) using apalladium or nickel catalyst in a reaction inert solvent such astoluene, 1,2-dimethoxyethane, dioxane, DMSO, DMF, or THF, in thepresence of a suitable ligand, and a base such as sodium, potassium, orlithium tert-butoxide, or cesium carbonate, at a temperature between 10°C. and 130° C. by the methods described in the literature such as: J.Med. Chem., 2007, 50, 2990-3003; Monatsh Chem, 2012, 143, 1575-1592; J.Med. Chem., 2011, 54, 6342-6363; Org. Proc. Res. Dev. 2014, 18,1145-1152 or other methods known to those skilled in the art.

Intermediate (4) may be prepared from ester (3a) via a hydrolysisreaction under conditions well known to those skilled in the art.Preferably, intermediate (3a, R=methyl or ethyl) is treated with anaqueous base such as sodium hydroxide, lithium hydroxide, or potassiumhydroxide, in a suitable solvent or solvent mixture comprised of water,methanol, and/or THF, at a temperature between 20° C. and 60° C.

Compounds of Formula (I) may be prepared from acid (4) and amine (5)under amide forming conditions well known to those skilled in the art,using coupling reagents such as propane phosphonic acid anhydride (T₃P),1,1′-carbonyldiimidazole (CDI),benzotriazo-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(BOP), 2-(1H-7-azabenzotriazol-1-yl)-1, 1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (HATU), oxalyl chloride,O-benzotriazol-1-yl-N,N,N′,N′-tetramethyluronium hexafluoro phosphate(HBTU), 2-chloro-1,3-dimethylimidazolinium chloride (DMC),N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide (EDCI) or1-hydroxybenzotriazole (HOBT) in a reaction inert solvent such asacetonitrile, dichloromethane (DCM), DMF, DMSO, or THF in the presenceof a base such as triethylamine, N-methyl-morpholine, orN,N-diisopropylethylamine at a temperature between 10° C. and 90° C.,preferably between 20° C. and 65° C.

Alternatively, compounds of Formula (I) may be prepared by a two-stepsequence from intermediate (1b) and amine (5) via an amide couplingreaction to afford intermediate (1c), followed by a metal mediatedcoupling reaction with aryl halide (2a). Preferably, intermediate (1c)is prepared from acid chloride (1b, Y═Cl) and amine (5) in the presenceof a base such as triethylamine or N,N-diisopropylethylamine, in areaction inert solvent, such as dichloromethane, at a temperaturebetween −20° C. to 30° C., preferably between −20° C. and 0° C.Alternatively, intermediate (1c) may be prepared from acid (1b, Y═OH)and amine (5) in the presence of an amide coupling reagent, such aspropane phosphonic acid anhydride (T₃P), 1,1′-carbonyldiimidazole (CDI),benzotriazo-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(BOP), 2-(1H-7-azabenzotriazol-1-yl)-1, 1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium (HATU),O-benzotriazol-1-yl-N,N,N′,N′-tetramethyluronium hexafluoro phosphate(HBTU), 2-chloro-1,3-dimethylimidazolinium chloride (DMC),N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide (EDCI) or1-hydroxybenzotriazole (HOBT) in a reaction inert solvent such asacetonitrile, dichloromethane, DMF, DMSO, or THF in the presence of abase such as triethylamine, N-methyl-morpholine, orN,N-diisopropylethylamine at a temperature between 10° C. and 90° C.Compounds of Formula (I) may then be prepared from halides (1c) and (2a)in a transition metal mediated coupling reaction. One of the halides(1c) or (2a) may be converted to an organometallic reagent, such as aboronic acid, zincate, stannane, or Grignard derivative using methodswell known to those skilled in the art. The resulting organometallicreagent may then be reacted with the other halide intermediate in atransition metal catalyzed cross coupling reaction. Preferably,intermediate (2a) is converted to a zincate and is coupled tointermediate (1c) using a palladium or nickel catalyst in a reactioninert solvent such as toluene, 1,2-dimethoxyethane, dioxane, DMSO, DMF,or THF, in the presence of a suitable ligand, and a base such as sodium,potassium, or lithium tert-butoxide, or cesium carbonate, at atemperature between 10° C. and 130° C. by the methods described in theliterature such as: J. Med. Chem., 2007, 50, 2990-3003; Monatsh Chem,2012, 143, 1575-1592; J. Med. Chem., 2011, 54, 6342-6363; Org. Proc.Res. Dev. 2014, 18, 1145-1152 or other methods known to those skilled inthe art.

Alternatively, compounds of Formula (I) may be prepared fromintermediate (1c) by a three-step sequence involving addition ofheteroaryl halide (2b) followed by demethylation and addition of arylhalide (6a). Intermediate (3b) may be prepared via transition metalmediated coupling reaction starting from halides (1c) and (2b). One ofthe halides (1c) or (2b) may be converted to an organometallic reagent,such as boronic acid, zincate, stannane, or Grignard derivatives usingmethods well known to those skilled in the art. The resultingorganometallic reagent may then be reacted with the other halideintermediate in a transition metal catalyzed cross coupling reaction.Preferably, intermediate (2b) is converted to a zincate and is coupledto intermediate (1c) using a palladium or nickel catalyst in a reactioninert solvent such as toluene, 1,2-dimethoxyethane, dioxane, DMSO, DMF,or THF, in the presence of a suitable ligand, and a base such as sodium,potassium, or lithium tert-butoxide, or cesium carbonate, at atemperature between 10° C. and 130° C. by the methods described in theliterature such as: J. Med. Chem., 2007, 50, 2990-3003; Monatsh Chem,2012, 143, 1575-1592; J. Med. Chem., 2011, 54, 6342-6363; Org. Proc.Res. Dev. 2014, 18, 1145-1152 or other methods known to those skilled inthe art. Intermediate (3c) may be prepared from intermediate (3b) viademethylation using hydrohalic acids such as hydrogen bromide, basessuch as sodium hydroxide or sodium alkoxide, boron tribromide, thiol orother methods known to those skilled in the art. For example,demethylation may be accomplished by methods described in the literaturesuch as: Arch Pharm Res 2008, 31, 305-309; Tetrahedron, 2005, 61,7833-7863; Protecting Groups in Organic Synthesis, John Wiley & Sons,Inc., Hoboken, N.J., USA, 2007, 370-382. Compounds of Formula (I) maythen be prepared from heteroaryl halide (6a) in a nucleophilic aromaticsubstitution reaction by alcohol (3c) in a reaction inert solvent suchas dimethylsulfoxide (DMSO), N,N-dimethylformamide (DMF), acetonitrile,or tetrahydrofuran (THF), in the presence of a suitable base, such ascesium carbonate, triethylamine (TEA) or N,N-diisopropylethylamine(DIPEA) at a temperature between 20° C. and 160° C. Preferably,intermediates (6a) and (3c) are reacted in DMSO, THF, or acetonitrile inthe presence of triethylamine or N,N-diisopropylethylamine, at atemperature between 100° C. and 160° C. to provide compounds of Formula(I) by methods described in the literature such as: Tetrahedron 2005, 626000-6005, Journal of Medicinal Chemistry, 2015, 58(7), 3036-3059.

Reaction Scheme II outlines the synthesis of intermediates (2a).

Intermediates (6a), (6b), and (2c) are commercially available or aredescribed in the literature and may be prepared via methods known tothose skilled in the art. Intermediate (2a) may be synthesized vianucleophilic aromatic substitution reaction of heteroaryl halide (6a)with hydroxypyridine (2c) in a reaction inert solvent such asdimethylsulfoxide (DMSO), N,N-dimethylformamide (DMF), acetonitrile,N-methyl-2-pyrrolidinone (NMP), or tetrahydrofuran (THF), in thepresence of a suitable base, such as cesium carbonate, potassiumcarbonate, triethylamine (TEA) or N,N-diisopropylethylamine (DIPEA) at atemperature between 20° C. and 160° C. Preferably, intermediates (6a)and (2c) are reacted in DMSO, NMP, or acetonitrile in the presence oftriethylamine or N,N-diisopropylethylamine, at a temperature between100° C. and 160° C. to provide intermediate (2a) using methods describedin the literature such as: Tetrahedron 2005, 62 6000-6005, Journal ofMedicinal Chemistry, 2015, 58(7), 3036-3059. Alternatively, intermediate(2a) may be synthesized by transition metal promoted ether formationbetween a hydroxy-aromatic coupling partner (2c) and an aromatic halide(6a) using methods such as those described in: Advanced Synthesis &Catalysis, 2011, 353, 3403-3414; Chemistry—A European Journal, 2015, 21,8727-8732; Synlett 2012, 23, 101; J. Org. Chem. 2009, 74, 7187; Org.Lett. 2007, 9, 643; Angew. Chem. Int. Ed. 2011, 50, 9943; J. Am. Chem.Soc. 2005, 127, 8146; J. Org. Chem. 2008, 73, 284; Org. Lett. 2002, 4,973. The appropriate starting materials (6a) and (2c) may be treatedwith a metal salt, such as copper(I) chloride, copper(I)bromide, orcopper(I) iodide, and a ligand such as2,2,6,6-tetramethylheptane-3,5-dione, 1,10-phenanthroline, or othersuitable ligand, in a reaction inert solvent such as toluene, DMSO, orDMF, in the presence of a base such as potassium carbonate, cesiumcarbonate, or potassium phosphate, at a temperature of 80° C. to 120° C.Preferably, the appropriate starting materials (6a) and (2c) are treatedwith copper(I) chloride and 2,2,6,6-tetramethylheptane-3,5-dione, intoluene, in the presence cesium carbonate, at a temperature of 100° C.to 120° C.

Alternatively, intermediate (2a) may be prepared from a two-stepsequence involving formation of N-oxide (6c) followed by addition ofhydroxyl pyridine (2c). N-oxide (6c) may be prepared from oxidizingagents such as m-chloroperoxybenzoic acid, hydrogen peroxide, potassiumpermanganate, or other oxidizing agents known to those skilled in theart in a reaction inert solvent such as dichloromethane,1,2-dichloroethane, or acetonitrile at a temperature between 0° C. and25° C. Preferably, intermediate (6b) is reacted in dichloromethane withm-chloroperoxybenzoic acid at a temperature between 10° C. and 25° C. toprovide intermediate (6c). Intermediate (2a) may be prepared fromintermediate (6c) and intermediate (2c) in the presence ofbromotripyrrolidinophosphonium hexafluorophosphate (PyBroP) in areaction inert solvent such as tetrahydrofuran, dichloromethane, ordioxane at a temperature between 10° C. and 25° C. Preferably,intermediate (6c) are reacted with intermediate (2c) in the presence ofbromotripyrrolidinophosphonium hexafluorophosphate in tetrahydrofuran ata temperature between 10° C. and 25° C. as described in Org. Lett.,2011, 13, 1840-1843.

Combination Agents

The compounds of the present invention can be administered alone or incombination with one or more additional therapeutic agents. By“administered in combination” or “combination therapy” it is meant thata compound of the present invention and one or more additionaltherapeutic agents are administered concurrently to the mammal beingtreated. When administered in combination each component may beadministered at the same time or sequentially in any order at differentpoints in time. Thus, each component may be administered separately butsufficiently closely in time so as to provide the desired therapeuticeffect. Thus, the methods of prevention and treatment described hereininclude use of combination agents.

The combination agents are administered to a mammal in a therapeuticallyeffective amount. By “therapeutically effective amount” it is meant anamount of a compound of the present invention that, when administeredalone or in combination with an additional therapeutic agent to amammal, is effective to treat the desired disease/condition e.g.,obesity, diabetes, and cardiovascular conditions such asanti-hypertensive agents and coronary heart disease.

Examples of suitable anti-diabetic agents include (e.g. insulins,metfomin, DPPIV inhibitors, GLP-1 agonists, analogues and mimetics,SGLT1 and SGLT2 inhibitors). Suitable anti-diabetic agents include anacetyl-CoA carboxylase-(ACC) inhibitor such as those described inWO2009144554, WO2003072197, WO2009144555 and WO2008065508, adiacylglycerol O-acyltransferase 1 (DGAT-1) inhibitor, such as thosedescribed in WO09016462 or WO2010086820, AZD7687 or LCQ908,monoacylglycerol O-acyltransferase inhibitors, a phosphodiesterase(PDE)-10 inhibitor, an AMPK activator, a sulfonylurea (e.g.,acetohexamide, chlorpropamide, diabinese, glibenclamide, glipizide,glyburide, glimepiride, gliclazide, glipentide, gliquidone, glisolamide,tolazamide, and tolbutamide), a meglitinide, an α-amylase inhibitor(e.g., tendamistat, trestatin and AL-3688), an α-glucoside hydrolaseinhibitor (e.g., acarbose), an α-glucosidase inhibitor (e.g., adiposine,camiglibose, emiglitate, miglitol, voglibose, pradimicin-Q, andsalbostatin), a PPARy agonist (e.g., balaglitazone, ciglitazone,darglitazone, englitazone, isaglitazone, pioglitazone androsiglitazone), a PPAR α/γ agonist (e.g., CLX-0940, GW-1536, GW-1929,GW-2433, KRP-297, L-796449, LR-90, MK-0767 and SB-219994), a biguanide(e.g., metformin), a glucagon-like peptide 1 (GLP-1) modulator such asan agonist (e.g., exendin-3 and exendin-4), liraglutide, albiglutide,exenatide (Byetta®), albiglutide, lixisenatide, dulaglutide,semaglutide, NN-9924, TTP-054, a protein tyrosine phosphatase-1B(PTP-1B) inhibitor (e.g., trodusquemine, hyrtiosal extract, andcompounds disclosed by Zhang, S., et al., Drug Discovery Today,12(9/10), 373-381 (2007)), SIRT-1 activator (e.g., resveratrol,GSK2245840 or GSK184072), a dipeptidyl peptidease IV (DPP-IV) inhibitor(e.g., those in WO2005116014, sitagliptin, vildagliptin, alogliptin,dutogliptin, linagliptin and saxagliptin), an insulin secreatagogue, afatty acid oxidation inhibitor, an A2 antagonist, a c-jun amino-terminalkinase (JNK) inhibitor, glucokinase activators (GKa) such as thosedescribed in WO2010103437, WO2010103438, WO2010013161, WO2007122482,TTP-399, TTP-355, TTP-547, AZD1656, ARRY403, MK-0599, TAK-329, AZD5658or GKM-001, insulin, an insulin mimetic, a glycogen phosphorylaseinhibitor (e.g. GSK1362885), a VPAC2 receptor agonist, SGLT2 inhibitors,such as those described in E. C. Chao et al. Nature Reviews DrugDiscovery 9, 551-559 (July 2010) including dapagliflozin, canagliflozin,empagliflozin, tofogliflozin (CSG452), Ertugliflozin, ASP-1941, THR1474,TS-071, ISIS388626 and LX4211 as well as those in WO2010023594, aglucagon receptor modulator such as those described in Demong, D. E. etal. Annual Reports in Medicinal Chemistry 2008, 43, 119-137, GPR119modulators, particularly agonists, such as those described inWO2010140092, WO2010128425, WO2010128414, WO2010106457, Jones, R. M. etal. in Medicinal Chemistry 2009, 44, 149-170 (e.g. MBX-2982, GSK1292263,APD597 and PSN821), FGF21 derivatives or analogs such as those describedin Kharitonenkov, A. et al. et al., Current Opinion in InvestigationalDrugs 2009, 10(4)359-364, TGR5 (also termed GPBAR1) receptor modulators,particularly agonists, such as those described in Zhong, M., CurrentTopics in Medicinal Chemistry, 2010, 10(4), 386-396 and INT777, GPR40agonists, such as those described in Medina, J. C., Annual Reports inMedicinal Chemistry, 2008, 43, 75-85, including but not limited toTAK-875, GPR120 modulators, particularly agonists, high affinitynicotinic acid receptor (HM74A) activators, and SGLT1 inhibitors, suchas GSK1614235. A further representative listing of anti-diabetic agentsthat can be combined with the compounds of the present invention can befound, for example, at page 28, line 35 through page 30, line 19 ofWO2011005611. Preferred anti-diabetic agents are metformin and DPP-IVinhibitors (e.g., sitagliptin, vildagliptin, alogliptin, dutogliptin,linagliptin and saxagliptin). Other antidiabetic agents could includeinhibitors or modulators of carnitine palmitoyl transferase enzymes,inhibitors of fructose 1,6-diphosphatase, inhibitors of aldosereductase, mineralocorticoid receptor inhibitors, inhibitors of TORC2,inhibitors of CCR2 and/or CCR5, inhibitors of PKC isoforms (e.g. PKCα,PKCβ, PKCγ), inhibitors of fatty acid synthetase, inhibitors of serinepalmitoyl transferase, modulators of GPR81, GPR39, GPR43, GPR41, GPR105,Kv1.3, retinol binding protein 4, glucocorticoid receptor, somatostainreceptors (e.g. SSTR1, SSTR2, SSTR3 and SSTR5), inhibitors or modulatorsof PDHK2 or PDHK4, inhibitors of MAP4K4, modulators of IL1 familyincluding ILlbeta, modulators of RXRalpha. In addition suitableanti-diabetic agents include mechanisms listed by Carpino, P. A.,Goodwin, B. Expert Opin. Ther. Pat, 2010, 20(12), 1627-51.

Suitable anti-obesity agents include 11(3-hydroxy steroiddehydrogenase-1 (11(3-HSD type 1) inhibitors, stearoyl-CoA desaturase-1(SCD-1) inhibitor, MCR-4 agonists, cholecystokinin-A (CCK-A) agonists,monoamine reuptake inhibitors (such as sibutramine), sympathomimeticagents, β₃ adrenergic agonists, dopamine agonists (such asbromocriptine), melanocyte-stimulating hormone analogs, 5HT2c agonists,melanin concentrating hormone antagonists, leptin (the OB protein),leptin analogs, leptin agonists, galanin antagonists, lipase inhibitors(such as tetrahydrolipstatin, i.e. orlistat), anorectic agents (such asa bombesin agonist), neuropeptide-Y antagonists (e.g., NPY Y5antagonists), PYY₃₋₃₆ (including analogs thereof), thyromimetic agents,dehydroepiandrosterone or an analog thereof, glucocorticoid agonists orantagonists, orexin antagonists, glucagon-like peptide-1 agonists,ciliary neurotrophic factors (such as Axokine™ available from RegeneronPharmaceuticals, Inc., Tarrytown, N.Y. and Procter & Gamble Company,Cincinnati, Ohio), human agouti-related protein (AGRP) inhibitors,ghrelin antagonists, histamine 3 antagonists or inverse agonists,neuromedin U agonists, MTP/ApoB inhibitors (e.g., gut-selective MTPinhibitors, such as dirlotapide), opioid antagonist, orexin antagonist,the combination of naltrexone with buproprion and the like.

Preferred anti-obesity agents for use in the combination aspects of thepresent invention include gut-selective MTP inhibitors (e.g.,dirlotapide, mitratapide and implitapide, R56918 (CAS No. 403987) andCAS No. 913541-47-6), CCKa agonists (e.g.,N-benzyl-2-[4-(1H-indol-3-ylmethyl)-5-oxo-1-phenyl-4,5-dihydro-2,3,6,10b-tetraaza-benzo[e]azulen-6-yl]-N-isopropyl-acetamidedescribed in PCT Publication No. WO 2005/116034 or US Publication No.2005-0267100 A1), 5HT2c agonists (e.g., lorcaserin), MCR4 agonist (e.g.,compounds described in U.S. Pat. No. 6,818,658), lipase inhibitor (e.g.,Cetilistat), PYY₃₋₃₆ (as used herein “PYY₃₋₃₆” includes analogs, such aspeglated PYY₃₋₃₆ e.g., those described in US Publication 2006/0178501),opioid antagonists (e.g., naltrexone), the combination of naltrexonewith buproprion, oleoyl-estrone (CAS No. 180003-17-2), obinepitide(TM30338), pramlintide (Symlin®), tesofensine (NS2330), leptin,liraglutide, bromocriptine, orlistat, exenatide (Byetta®), AOD-9604 (CASNo. 221231-10-3), phentermine and topiramate (trade name: Qsymia), andsibutramine. Preferably, compounds of the present invention andcombination therapies are administered in conjunction with exercise anda sensible diet.

The compounds of the present invention may be used in combination withcholesterol modulating agents (including cholesterol lowering agents)such as a lipase inhibitor, an HMG-CoA reductase inhibitor, an HMG-CoAsynthase inhibitor, an HMG-CoA reductase gene expression inhibitor, anHMG-CoA synthase gene expression inhibitor, an MTP/Apo B secretioninhibitor, a CETP inhibitor, a bile acid absorption inhibitor, acholesterol absorption inhibitor, a cholesterol synthesis inhibitor, asqualene synthetase inhibitor, a squalene epoxidase inhibitor, asqualene cyclase inhibitor, a combined squalene epoxidase/squalenecyclase inhibitor, a fibrate, niacin, an ion-exchange resin, anantioxidant, an ACAT inhibitor or a bile acid sequestrant or an agentsuch as mipomersen.

Examples of suitable cholesterol/lipid lowering agents and lipid profiletherapies include: HMG-CoA reductase inhibitors (e.g., pravastatin,lovastatin, atorvastatin, simvastatin, fluvastatin, NK-104 (a.k.a.itavastatin, or nisvastatin or nisbastatin) and ZD-4522 (a.k.a.rosuvastatin, or atavastatin or visastatin); squalene synthetaseinhibitors; fibrates; bile acid sequestrants (such as questran); ACATinhibitors; MTP inhibitors; lipooxygenase inhibitors; choesterolabsorption inhibitors; and cholesteryl ester transfer proteininhibitors. Other atherosclerotic agents include PCSK9 modulators.

In another embodiment, a compound of Formula I may be co-administeredwith agents for the treatment of non-alcoholic steatohepatitis (NASH)and/or non-alcoholic fatty liver disease (NAFLD), such as Orlistat, TZDsand other insulin sensitizing agents, FGF21 analogs, Metformin,Omega-3-acid ethyl esters (e.g. Lovaza), Fibrates, HMG CoA-reductaseInhibitors, Ezitimbe, Probucol, Ursodeoxycholic acid, TGR5 agonists, FXRagonists, Vitamin E, Betaine, Pentoxifylline, CB1 antagonists,Carnitine, N-acetylcysteine, Reduced glutathione, lorcaserin, thecombination of naltrexone with buproprion, SGLT2 Inhibitors,Phentermine, Topiramate, Incretin (GLP and GIP) analogs andAngiotensin-receptor blockers.

In another embodiment, the additional pharmaceutical agent is selectedfrom the group consisting of cysteamine or a pharmaceutically acceptablesalt thereof, cystamine or a pharmaceutically acceptable salt thereof,an anti-oxidant compound, lecithin, vitamin B complex, a bile saltpreparations, an antagonists of Cannabinoid-1 (CB1) receptor, an inverseagonists of Cannabinoid-1 (CB1) receptor, a peroxisomeproliferator-activated receptor) activity regulators, a benzothiazepineor benzothiepine compound, an RNA antisense construct to inhibit proteintyrosine phosphatase PTPRU, a heteroatom-linked substituted piperidineand derivatives thereof, an azacyclopentane derivative capable ofinhibiting stearoyl-coenzyme alpha delta-9 desaturase, acylamidecompound having secretagogue or inducer activity of adiponectin, aquaternary ammonium compound, Glatiramer acetate, pentraxin proteins, aHMG-CoA reductase inhibitor, n-acetyl cysteine, isoflavone compound, amacrolide antibiotic, a galectin inhibitor, an antibody, or anycombination of thereof.

Additional therapeutic agents include anti-coagulant or coagulationinhibitory agents, anti-platelet or platelet inhibitory agents, thrombininhibitors, thrombolytic or fibrinolytic agents, anti-arrythmic agents,anti-hypertensive agents, calcium channel blockers (L-type and T-type),cardiac glycosides, diruetics, mineralocorticoid receptor antagonists,NO donating agents such as organonitrates, NO promoting agents such asphosphodiesterase inhibitors, cholesterol/lipid lowering agents andlipid profile therapies, anti-diabetic agents, anti-depressants,anti-inflammatory agents (steroidal and non-steroidal),anti-osteoporosis agents, hormone replacement therapies, oralcontraceptives, anti-obesity agents, anti-anxiety agents,anti-proliferative agents, anti-tumor agents, anti-ulcer andgastroesophageal reflux disease agents, growth hormone and/or growthhormone secretagogues, thyroid mimetics (including thyroid hormonereceptor antagonist), anti-infective agents, anti-viral agents,anti-bacterial agents, and anti-fungal agents.

Agents used in an ICU setting are included, for example, dobutamine,dopamine, dpinephrine, nitroglycerin, nitroprusside etc.

Combination agents useful for treating vasculitis are included, forexample, azathioprine, cyclophosphamide, mycophenolate, mofetil,rituximab etc.

In another embodiment, the present invention provides a combinationwherein the second agent is at least one agent selected from a factor Xainhibitor, an anti-coagulant agent, an anti-platelet agent, a thrombininhibiting agent, a thrombolytic agent, and a fibrinolytic agent.Exemplary factor Xa inhibitors include apixaban and rivaroxaban.Examples of suitable anti-coagulants for use in combination with thecompounds of the present invention include heparins (e.g., unfractionedand low molecular weight heparins such as enoxaparin and dalteparin).

In another preferred embodiment the second agent is at least one agentselected from warfarin, dabigatran, unfractionated heparin, lowmolecular weight heparin, synthetic pentasaccharide, hirudin,argatrobanas, aspirin, ibuprofen, naproxen, sulindac, indomethacin,mefenamate, droxicam, diclofenac, sulfinpyrazone, piroxicam,ticlopidine, clopidogrel, tirofiban, eptifibatide, abciximab,melagatran, disulfatohirudin, tissue plasminogen activator, modifiedtissue plasminogen activator, anistreplase, urokinase, andstreptokinase.

A preferred second agent is at least one anti-platelet agent. Especiallypreferred anti-platelet agents are aspirin and clopidogrel.

The term anti-platelet agents (or platelet inhibitory agents), as usedherein, denotes agents that inhibit platelet function, for example byinhibiting the aggregation, adhesion or granular secretion of platelets.Agents include, but are not limited to, the various known non-steroidalanti-inflammatory drugs (NSAIDS) such as aspirin, ibuprofen, naproxen,sulindac, indomethacin, mefenamate, droxicam, diclofenac,sulfinpyrazone, piroxicam, and pharmaceutically acceptable salts orprodrugs thereof. Of the NSAIDS, aspirin (acetylsalicyclic acid or ASA)and COX-2 inhibitors such as CELEBREX or piroxicam are preferred. Othersuitable platelet inhibitory agents include IIb/IIIa antagonists (e.g.,tirofiban, eptifibatide, and abciximab), thromboxane-A2-receptorantagonists (e.g., ifetroban), thromboxane-A2-synthetase inhibitors,PDE-III inhibitors (e.g., Pletal, dipyridamole), and pharmaceuticallyacceptable salts or prodrugs thereof.

The term anti-platelet agents (or platelet inhibitory agents), as usedherein, is also intended to include ADP (adenosine diphosphate) receptorantagonists, preferably antagonists of the purinergic receptors P₂Y₁ andP₂Y₁₂, with P₂Y₁₂ being even more preferred. Preferred P₂Y₁₂ receptorantagonists include ticagrelor, prasugrel, ticlopidine and clopidogrel,including pharmaceutically acceptable salts or prodrugs thereof.Clopidogrel is an even more preferred agent. Ticlopidine and clopidogrelare also preferred compounds since they are known to be gentle on thegastro-intestinal tract in use.

The term thrombin inhibitors (or anti-thrombin agents), as used herein,denotes inhibitors of the serine protease thrombin. By inhibitingthrombin, various thrombin-mediated processes, such as thrombin-mediatedplatelet activation (that is, for example, the aggregation of platelets,and/or the granular secretion of plasminogen activator inhibitor-1and/or serotonin) and/or fibrin formation are disrupted. A number ofthrombin inhibitors are known to one of skill in the art and theseinhibitors are contemplated to be used in combination with the presentcompounds. Such inhibitors include, but are not limited to, boroargininederivatives, boropeptides, dabigatran, heparins, hirudin, argatroban,and melagatran, including pharmaceutically acceptable salts and prodrugsthereof. Boroarginine derivatives and boropeptides include N-acetyl andpeptide derivatives of boronic acid, such as C-terminalalpha-aminoboronic acid derivatives of lysine, ornithine, arginine,homoarginine and corresponding isothiouronium analogs thereof. The termhirudin, as used herein, includes suitable derivatives or analogs ofhirudin, referred to herein as hirulogs, such as disulfatohirudin. Theterm thrombolytics or fibrinolytic agents (or thrombolytics orfibrinolytics), as used herein, denote agents that lyse blood clots(thrombi). Such agents include tissue plasminogen activator (natural orrecombinant) and modified forms thereof, anistreplase, urokinase,streptokinase, tenecteplase (TNK), lanoteplase (nPA), factor VIIainhibitors, PAI-1 inhibitors (i.e., inactivators of tissue plasminogenactivator inhibitors), alpha2-antiplasmin inhibitors, and anisoylatedplasminogen streptokinase activator complex, including pharmaceuticallyacceptable salts or prodrugs thereof. The term anistreplase, as usedherein, refers to anisoylated plasminogen streptokinase activatorcomplex, as described, for example, in EP 028,489, the disclosure ofwhich is hereby incorporated herein by reference herein. The termurokinase, as used herein, is intended to denote both dual and singlechain urokinase, the latter also being referred to herein asprourokinase.

Examples of suitable anti-arrythmic agents include: Class I agents (suchas propafenone); Class II agents (such as metoprolol, atenolol,carvadiol and propranolol); Class III agents (such as sotalol,dofetilide, amiodarone, azimilide and ibutilide); Class IV agents (suchas ditiazem and verapamil); K⁺ channel openers such as IAch inhibitors,and I_(Kur) inhibitors (e.g., compounds such as those disclosed inWO01/40231).

The compounds of the present invention may be used in combination withantihypertensive agents and such antihypertensive activity is readilydetermined by those skilled in the art according to standard assays(e.g., blood pressure measurements).

Examples of suitable anti-hypertensive agents include: alpha adrenergicblockers; beta adrenergic blockers; calcium channel blockers (e.g.,diltiazem, verapamil, nifedipine and amlodipine); vasodilators (e.g.,hydralazine), diruetics (e.g., chlorothiazide, hydrochlorothiazide,flumethiazide, hydroflumethiazide, bendroflumethiazide,methylchlorothiazide, trichloromethiazide, polythiazide, benzthiazide,ethacrynic acid tricrynafen, chlorthalidone, torsemide, furosemide,musolimine, bumetanide, triamtrenene, amiloride, spironolactone); renininhibitors; ACE inhibitors (e.g., captopril, zofenopril, fosinopril,enalapril, ceranopril, cilazopril, delapril, pentopril, quinapril,ramipril, lisinopril); AT-1 receptor antagonists (e.g., losartan,irbesartan, valsartan); ET receptor antagonists (e.g., sitaxsentan,atrsentan and compounds disclosed in U.S. Pat. Nos. 5,612,359 and6,043,265); Dual ET/AII antagonist (e.g., compounds disclosed in WO00/01389); neutral endopeptidase (NEP) inhibitors; vasopepsidaseinhibitors (dual NEP-ACE inhibitors) (e.g., gemopatrilat and nitrates).An exemplary antianginal agent is ivabradine.

Examples of suitable calcium channel blockers (L-type or T-type) includediltiazem, verapamil, nifedipine and amlodipine and mybefradil.

Examples of suitable cardiac glycosides include digitalis and ouabain.

In one embodiment, a Formula I compound may be co-administered with oneor more diuretics. Examples of suitable diuretics include (a) loopdiuretics such as furosemide (such as LASIX™), torsemide (such asDEMADEX™), bemetanide (such as BUMEX™), and ethacrynic acid (such asEDECRIN™); (b) thiazide-type diuretics such as chlorothiazide (such asDIURIL™, ESIDRIX™ or HYDRODIURIL™), hydrochlorothiazide (such asMICROZIDE™ or ORETIC™), benzthiazide, hydroflumethiazide (such asSALURON™), bendroflumethiazide, methychlorthiazide, polythiazide,trichlormethiazide, and indapamide (such as LOZOL™); (c)phthalimidine-type diuretics such as chlorthalidone (such as HYGROTON™),and metolazone (such as ZAROXOLYN™); (d) quinazoline-type diuretics suchas quinethazone; and (e) potassium-sparing diuretics such as triamterene(such as DYRENIUM™), and amiloride (such as MIDAMOR™ or MODURETIC™).

In another embodiment, a compound of Formula I may be co-administeredwith a loop diuretic. In still another embodiment, the loop diuretic isselected from furosemide and torsemide. In still another embodiment, oneor more compounds of Formula I or Ia may be co-administered withfurosemide. In still another embodiment, one or more compounds ofFormula I or Ia may be co-administered with torsemide which mayoptionally be a controlled or modified release form of torsemide.

In another embodiment, a compound of Formula I may be co-administeredwith a thiazide-type diuretic. In still another embodiment, thethiazide-type diuretic is selected from the group consisting ofchlorothiazide and hydrochlorothiazide. In still another embodiment, oneor more compounds of Formula I or Ia may be co-administered withchlorothiazide. In still another embodiment, one or more compounds ofFormula I or Ia may be co-administered with hydrochlorothiazide.

In another embodiment, one or more compounds of Formula I or Ia may beco-administered with a phthalimidine-type diuretic. In still anotherembodiment, the phthalimidine-type diuretic is chlorthalidone. Examplesof suitable mineralocorticoid receptor antagonists includesprionolactone and eplerenone. Examples of suitable phosphodiesteraseinhibitors include: PDE III inhibitors (such as cilostazol); and PDE Vinhibitors (such as sildenafil).

Those skilled in the art will recognize that the compounds of thisinvention may also be used in conjunction with other cardiovascular orcerebrovascular treatments including PCI, stenting, drug eluting stents,stem cell therapy and medical devices such as implanted pacemakers,defibrillators, or cardiac resynchronization therapy.

The dosage of the additional pharmaceutical agent is generally dependentupon a number of factors including the health of the subject beingtreated, the extent of treatment desired, the nature and kind ofconcurrent therapy, if any, and the frequency of treatment and thenature of the effect desired. In general, the dosage range of theadditional pharmaceutical agent is in the range of from about 0.001 mgto about 100 mg per kilogram body weight of the individual per day,preferably from about 0.1 mg to about 10 mg per kilogram body weight ofthe individual per day. However, some variability in the general dosagerange may also be required depending upon the age and weight of thesubject being treated, the intended route of administration, theparticular anti-obesity agent being administered and the like. Thedetermination of dosage ranges and optimal dosages for a particularpatient is also well within the ability of one of ordinary skill in theart having the benefit of the instant disclosure.

According to the methods of treatment of the invention, a compound ofthe present invention or a combination of a compound of the presentinvention and at least one additional pharmaceutical agent (referred toherein as a “combination”) is administered to a subject in need of suchtreatment, preferably in the form of a pharmaceutical composition. Inthe combination aspect of the invention, the compound of the presentinvention and at least one other pharmaceutical agent (e.g., anotheranti-obesity agent,) may be administered either separately or in apharmaceutical composition comprising both. It is generally preferredthat such administration be oral.

When a combination of a compound of the present invention and at leastone other pharmaceutical agent are administered together, suchadministration may be sequential in time or simultaneous. Simultaneousadministration of drug combinations is generally preferred. Forsequential administration, a compound of the present invention and theadditional pharmaceutical agent may be administered in any order. It isgenerally preferred that such administration be oral. It is especiallypreferred that such administration be oral and simultaneous. When acompound of the present invention and the additional pharmaceuticalagent are administered sequentially, the administration of each may beby the same or by different methods.

According to the methods of the invention, a compound of the presentinvention or a combination is preferably administered in the form of apharmaceutical composition. Accordingly, a compound of the presentinvention or a combination can be administered to a patient separatelyor together in any conventional oral, rectal, transdermal, parenteral(e.g., intravenous, intramuscular or subcutaneous), intracisternal,intravaginal, intraperitoneal, topical (e.g., powder, ointment, cream,spray or lotion), buccal or nasal dosage form (e.g., spray, drops orinhalant).

The compounds of the invention or combinations can be administered alonebut will generally be administered in an admixture with one or moresuitable pharmaceutical excipients, adjuvants, diluents or carriersknown in the art and selected with regard to the intended route ofadministration and standard pharmaceutical practice. The compound of theinvention or combination may be formulated to provide immediate-,delayed-, modified-, sustained-, pulsed- or controlled-release dosageforms depending on the desired route of administration and thespecificity of release profile, commensurate with therapeutic needs.

The pharmaceutical composition comprises a compound of the invention ora combination in an amount generally in the range of from about 1% toabout 75%, 80%, 85%, 90% or even 95% (by weight) of the composition,usually in the range of about 1%, 2% or 3% to about 50%, 60% or 70%,more frequently in the range of about 1%, 2% or 3% to less than 50% suchas about 25%, 30% or 35%.

Methods of preparing various pharmaceutical compositions with a specificamount of active compound are known to those skilled in this art. Forexamples, see Remington: The Practice of Pharmacy, Lippincott Williamsand Wilkins, Baltimore Md. 20.sup.th ed. 2000.

Compositions suitable for parenteral injection generally includepharmaceutically acceptable sterile aqueous or nonaqueous solutions,dispersions, suspensions, or emulsions, and sterile powders forreconstitution into sterile injectable solutions or dispersions.Examples of suitable aqueous and nonaqueous carriers or diluents(including solvents and vehicles) include water, ethanol, polyols(propylene glycol, polyethylene glycol, glycerol, and the like),suitable mixtures thereof, triglycerides including vegetable oils suchas olive oil, and injectable organic esters such as ethyl oleate. Apreferred carrier is Miglyol® brand caprylic/capric acid ester withglycerine or propylene glycol (e.g., Miglyol® 812, Miglyol® 829,Miglyol® 840) available from Condea Vista Co., Cranford, N.J. Properfluidity can be maintained, for example, by the use of a coating such aslecithin, by the maintenance of the required particle size in the caseof dispersions, and by the use of surfactants.

These compositions for parenteral injection may also contain excipientssuch as preserving, wetting, emulsifying, and dispersing agents.Prevention of microorganism contamination of the compositions can beaccomplished with various antibacterial and antifungal agents, forexample, parabens, chlorobutanol, phenol, sorbic acid, and the like. Itmay also be desirable to include isotonic agents, for example, sugars,sodium chloride, and the like. Prolonged absorption of injectablepharmaceutical compositions can be brought about by the use of agentscapable of delaying absorption, for example, aluminum monostearate andgelatin.

Solid dosage forms for oral administration include capsules, tablets,chews, lozenges, pills, powders, and multi-particulate preparations(granules). In such solid dosage forms, a compound of the presentinvention or a combination is admixed with at least one inert excipient,diluent or carrier. Suitable excipients, diluents or carriers includematerials such as sodium citrate or dicalcium phosphate and/or (a) oneor more fillers or extenders (e.g., microcrystalline cellulose(available as Avicel™ from FMC Corp.) starches, lactose, sucrose,mannitol, silicic acid, xylitol, sorbitol, dextrose, calcium hydrogenphosphate, dextrin, alpha-cyclodextrin, beta-cyclodextrin, polyethyleneglycol, medium chain fatty acids, titanium oxide, magnesium oxide,aluminum oxide and the like); (b) one or more binders (e.g.,carboxymethylcellulose, methylcellulose, hydroxypropylcellulose,hydroxypropylmethylcellulose, gelatin, gum arabic, ethyl cellulose,polyvinyl alcohol, pullulan, pregelatinized starch, agar, tragacanth,alginates, gelatin, polyvinylpyrrolidone, sucrose, acacia and the like);(c) one or more humectants (e.g., glycerol and the like); (d) one ormore disintegrating agents (e.g., agar-agar, calcium carbonate, potatoor tapioca starch, alginic acid, certain complex silicates, sodiumcarbonate, sodium lauryl sulphate, sodium starch glycolate (available asExplotab™ from Edward Mendell Co.), cross-linked polyvinyl pyrrolidone,croscarmellose sodium A-type (available as Ac-di-sol™), polyacrilinpotassium (an ion exchange resin) and the like); (e) one or moresolution retarders (e.g., paraffin and the like); (f) one or moreabsorption accelerators (e.g., quaternary ammonium compounds and thelike); (g) one or more wetting agents (e.g., cetyl alcohol, glycerolmonostearate and the like); (h) one or more adsorbents (e.g., kaolin,bentonite and the like); and/or (i)one or more lubricants (e.g., talc,calcium stearate, magnesium stearate, stearic acid, polyoxyl stearate,cetanol, talc, hydrogenated caster oil, sucrose esters of fatty acid,dimethylpolysiloxane, microcrystalline wax, yellow beeswax, whitebeeswax, solid polyethylene glycols, sodium lauryl sulfate and thelike). In the case of capsules and tablets, the dosage forms may alsocomprise buffering agents.

Solid compositions of a similar type may also be used as fillers in softor hard filled gelatin capsules using such excipients as lactose or milksugar, as well as high molecular weight polyethylene glycols, and thelike.

Solid dosage forms such as tablets, dragees, capsules, and granules maybe prepared with coatings and shells, such as enteric coatings andothers well known in the art. They may also contain opacifying agents,and can also be of such composition that they release the compound ofthe present invention and/or the additional pharmaceutical agent in adelayed manner. Examples of embedding compositions that can be used arepolymeric substances and waxes. The drug may also be inmicro-encapsulated form, if appropriate, with one or more of theabove-mentioned excipients.

For tablets, the active agent will typically comprise less than 50% (byweight) of the formulation, for example less than about 10% such as 5%or 2.5% by weight. The predominant portion of the formulation comprisesfillers, diluents, disintegrants, lubricants and optionally, flavors.The composition of these excipients is well known in the art.Frequently, the fillers/diluents will comprise mixtures of two or moreof the following components: microcrystalline cellulose, mannitol,lactose (all types), starch, and di-calcium phosphate. Thefiller/diluent mixtures typically comprise less than 98% of theformulation and preferably less than 95%, for example 93.5%. Preferreddisintegrants include Ac-di-sol™, Explotab™, starch and sodium laurylsulphate. When present a disintegrant will usually comprise less than10% of the formulation or less than 5%, for example about 3%. Apreferred lubricant is magnesium stearate. When present a lubricant willusually comprise less than 5% of the formulation or less than 3%, forexample about 1%.

Tablets may be manufactured by standard tabletting processes, forexample, direct compression or a wet, dry or melt granulation, meltcongealing process and extrusion. The tablet cores may be mono ormulti-layer(s) and can be coated with appropriate overcoats known in theart.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, solutions, suspensions, syrups, and elixirs. Inaddition to the compound of the present invention or the combination,the liquid dosage form may contain inert diluents commonly used in theart, such as water or other solvents, solubilizing agents andemulsifiers, as for example, ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformamide, oils (e.g., cottonseedoil, groundnut oil, corn germ oil, olive oil, castor oil, sesame seedoil and the like), Miglyole® (available from CONDEA Vista Co., Cranford,N.J.), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols andfatty acid esters of sorbitan, or mixtures of these substances, and thelike.

Besides such inert diluents, the composition may also includeexcipients, such as wetting agents, emulsifying and suspending agents,sweetening, flavoring, and perfuming agents.

Oral liquid forms of the compounds of the invention or combinationsinclude solutions, wherein the active compound is fully dissolved.Examples of solvents include all pharmaceutically precedented solventssuitable for oral administration, particularly those in which thecompounds of the invention show good solubility, e.g., polyethyleneglycol, polypropylene glycol, edible oils and glyceryl- andglyceride-based systems. Glyceryl- and glyceride-based systems mayinclude, for example, the following branded products (and correspondinggeneric products): Captex™ 355 EP (glyceryl tricaprylate/caprate, fromAbitec, Columbus Ohio), Crodamol™ GTC/C (medium chain triglyceride, fromCroda, Cowick Hall, UK) or Labrafac™ CC (medium chain triglyides, fromGattefosse), Captex™ 500P (glyceryl triacetate i.e. triacetin, fromAbitec), Capmul™ MCM (medium chain mono- and diglycerides, fromAbitec),Migyol™ 812 (caprylic/capric triglyceride, from Condea, Cranford N.J.),Migyol™ 829 (caprylic/capric/succinic triglyceride, from Condea),Migyol™ 840 (propylene glycol dicaprylate/dicaprate, from Condea),Labrafil™ M1944CS (oleoyl macrogol-6 glycerides, from Gattefosse),Peceol™ (glyceryl monooleate, from Gattefosse) and Maisine™ 35-1(glyceryl monooleate, from Gattefosse). Of particular interest are themedium chain (about C.sub.8 to C.sub.10) triglyceride oils. Thesesolvents frequently make up the predominant portion of the composition,i.e., greater than about 50%, usually greater than about 80%, forexample about 95% or 99%. Adjuvants and additives may also be includedwith the solvents principally as taste-mask agents, palatability andflavoring agents, antioxidants, stabilizers, texture and viscositymodifiers and solubilizers.

Suspensions, in addition to the compound of the present invention or thecombination, may further comprise carriers such as suspending agents,e.g., ethoxylated isostearyl alcohols, polyoxyethylene sorbitol andsorbitan esters, microcrystalline cellulose, aluminum metahydroxide,bentonite, agar-agar, and tragacanth, or mixtures of these substances,and the like.

Compositions for rectal or vaginal administration preferably comprisesuppositories, which can be prepared by mixing a compound of the presentinvention or a combination with suitable non-irritating excipients orcarriers, such as cocoa butter, polyethylene glycol or a suppository waxwhich are solid at ordinary room temperature, but liquid at bodytemperature, and therefore, melt in the rectum or vaginal cavity therebyreleasing the active component(s).

Dosage forms for topical administration of the compounds of the presentinvention or combinations include ointments, creams, lotions, powdersand sprays. The drugs are admixed with a pharmaceutically acceptableexcipient, diluent or carrier, and any preservatives, buffers, orpropellants that may be required.

Many of the present compounds are poorly soluble in water, e.g., lessthan about 1 .mu.g/mL. Therefore, liquid compositions in solubilizing,non-aqueous solvents such as the medium chain triglyceride oilsdiscussed above are a preferred dosage form for these compounds.

Solid amorphous dispersions, including dispersions formed by aspray-drying process, are also a preferred dosage form for the poorlysoluble compounds of the invention. By “solid amorphous dispersion” ismeant a solid material in which at least a portion of the poorly solublecompound is in the amorphous form and dispersed in a water-solublepolymer. By “amorphous” is meant that the poorly soluble compound is notcrystalline. By “crystalline” is meant that the compound exhibitslong-range order in three dimensions of at least 100 repeat units ineach dimension. Thus, the term amorphous is intended to include not onlymaterial which has essentially no order, but also material which mayhave some small degree of order, but the order is in less than threedimensions and/or is only over short distances. Amorphous material maybe characterized by techniques known in the art such as powder x-raydiffraction (PXRD) crystallography, solid state NMR, or thermaltechniques such as differential scanning calorimetry (DSC).

Preferably, at least a major portion (i.e., at least about 60 wt %) ofthe poorly soluble compound in the solid amorphous dispersion isamorphous. The compound can exist within the solid amorphous dispersionin relatively pure amorphous domains or regions, as a solid solution ofthe compound homogeneously distributed throughout the polymer or anycombination of these states or those states that lie intermediatebetween them. Preferably, the solid amorphous dispersion issubstantially homogeneous so that the amorphous compound is dispersed ashomogeneously as possible throughout the polymer. As used herein,“substantially homogeneous” means that the fraction of the compound thatis present in relatively pure amorphous domains or regions within thesolid amorphous dispersion is relatively small, on the order of lessthan 20 wt %, and preferably less than 10 wt % of the total amount ofdrug.

Water-soluble polymers suitable for use in the solid amorphousdispersions should be inert, in the sense that they do not chemicallyreact with the poorly soluble compound in an adverse manner, arepharmaceutically acceptable, and have at least some solubility inaqueous solution at physiologically relevant pHs (e.g. 1-8). The polymercan be neutral or ionizable, and should have an aqueous-solubility of atleast 0.1 mg/mL over at least a portion of the pH range of 1-8.

Water-soluble polymers suitable for use with the present invention maybe cellulosic or non-cellulosic. The polymers may be neutral orionizable in aqueous solution. Of these, ionizable and cellulosicpolymers are preferred, with ionizable cellulosic polymers being morepreferred.

Exemplary water-soluble polymers include hydroxypropyl methyl celluloseacetate succinate (HPMCAS), hydroxypropyl methyl cellulose (HPMC),hydroxypropyl methyl cellulose phthalate (HPMCP), carboxy methyl ethylcellulose (CMEC), cellulose acetate phthalate (CAP), cellulose acetatetrimellitate (CAT), polyvinylpyrrolidone (PVP), hydroxypropyl cellulose(HPC), methyl cellulose (MC), block copolymers of ethylene oxide andpropylene oxide (PEO/PPO, also known as poloxamers), and mixturesthereof. Especially preferred polymers include HPMCAS, HPMC, HPMCP,CMEC, CAP, CAT, PVP, poloxamers, and mixtures thereof. Most preferred isHPMCAS. See European Patent Application Publication No. 0 901 786 A2,the disclosure of which is incorporated herein by reference.

The solid amorphous dispersions may be prepared according to any processfor forming solid amorphous dispersions that results in at least a majorportion (at least 60%) of the poorly soluble compound being in theamorphous state. Such processes include mechanical, thermal and solventprocesses. Exemplary mechanical processes include milling and extrusion;melt processes including high temperature fusion, solvent-modifiedfusion and melt-congeal processes; and solvent processes includingnon-solvent precipitation, spray coating and spray drying. See, forexample, the following U.S. Patents, the pertinent disclosures of whichare incorporated herein by reference: U.S. Pat. Nos. 5,456,923 and5,939,099, which describe forming dispersions by extrusion processes;U.S. Pat. Nos. 5,340,591 and 4,673,564, which describe formingdispersions by milling processes; and U.S. Pat. Nos. 5,707,646 and4,894,235, which describe forming dispersions by melt congeal processes.In a preferred process, the solid amorphous dispersion is formed byspray drying, as disclosed in European Patent Application PublicationNo. 0 901 786 A2. In this process, the compound and polymer aredissolved in a solvent, such as acetone or methanol, and the solvent isthen rapidly removed from the solution by spray drying to form the solidamorphous dispersion. The solid amorphous dispersions may be prepared tocontain up to about 99 wt % of the compound, e.g., 1 wt %, 5 wt %, 10 wt%, 25 wt %, 50 wt %, 75 wt %, 95 wt %, or 98 wt % as desired.

The solid dispersion may be used as the dosage form itself or it mayserve as a manufacturing-use-product (MUP) in the preparation of otherdosage forms such as capsules, tablets, solutions or suspensions. Anexample of an aqueous suspension is an aqueous suspension of a 1:1 (w/w)compound/HPMCAS-HF spray-dried dispersion containing 2.5 mg/mL ofcompound in 2% polysorbate-80. Solid dispersions for use in a tablet orcapsule will generally be mixed with other excipients or adjuvantstypically found in such dosage forms. For example, an exemplary fillerfor capsules contains a 2:1 (w/w) compound/HPMCAS-MF spray-drieddispersion (60%), lactose (fast flow) (15%), microcrystalline cellulose(e.g., Avicel.sup.(R0-102) (15.8%), sodium starch (7%), sodium laurylsulfate (2%) and magnesium stearate (1%).

The HPMCAS polymers are available in low, medium and high grades asAqoa.sup.(R)-LF, Aqoat.sup.(R)-MF and Aqoat.sup.(R)-HF respectively fromShin-Etsu Chemical Co., LTD, Tokyo, Japan. The higher MF and HF gradesare generally preferred.

The following paragraphs describe exemplary formulations, dosages, etc.useful for non-human animals. The administration of the compounds of thepresent invention and combinations of the compounds of the presentinvention with anti-obesity agents can be effected orally or non-orally.

An amount of a compound of the present invention or combination of acompound of the present invention with another anti-obesity agent isadministered such that an effective dose is received. Generally, a dailydose that is administered orally to an animal is between about 0.01 andabout 1,000 mg/kg of body weight, e.g., between about 0.01 and about 300mg/kg or between about 0.01 and about 100 mg/kg or between about 0.01and about 50 mg/kg of body weight, or between about 0.01 and about 25mg/kg, or about 0.01 and about 10 mg/kg or about 0.01 and about 5 mg/kg.

Conveniently, a compound of the present invention (or combination) canbe carried in the drinking water so that a therapeutic dosage of thecompound is ingested with the daily water supply. The compound can bedirectly metered into drinking water, preferably in the form of aliquid, water-soluble concentrate (such as an aqueous solution of awater-soluble salt).

Conveniently, a compound of the present invention (or combination) canalso be added directly to the feed, as such, or in the form of an animalfeed supplement, also referred to as a premix or concentrate. A premixor concentrate of the compound in an excipient, diluent or carrier ismore commonly employed for the inclusion of the agent in the feed.Suitable excipients, diluents or carriers are liquid or solid, asdesired, such as water, various meals such as alfalfa meal, soybeanmeal, cottonseed oil meal, linseed oil meal, corncob meal and corn meal,molasses, urea, bone meal, and mineral mixes such as are commonlyemployed in poultry feeds. A particularly effective excipient, diluentor carrier is the respective animal feed itself; that is, a smallportion of such feed. The carrier facilitates uniform distribution ofthe compound in the finished feed with which the premix is blended.

Preferably, the compound is thoroughly blended into the premix and,subsequently, the feed. In this respect, the compound may be dispersedor dissolved in a suitable oily vehicle such as soybean oil, corn oil,cottonseed oil, and the like, or in a volatile organic solvent and thenblended with the carrier. It will be appreciated that the proportions ofcompound in the concentrate are capable of wide variation since theamount of the compound in the finished feed may be adjusted by blendingthe appropriate proportion of premix with the feed to obtain a desiredlevel of compound.

High potency concentrates may be blended by the feed manufacturer withproteinaceous carrier such as soybean oil meal and other meals, asdescribed above, to produce concentrated supplements, which are suitablefor direct feeding to animals. In such instances, the animals arepermitted to consume the usual diet. Alternatively, such concentratedsupplements may be added directly to the feed to produce a nutritionallybalanced, finished feed containing a therapeutically effective level ofa compound of the present invention. The mixtures are thoroughly blendedby standard procedures, such as in a twin shell blender, to ensurehomogeneity.

If the supplement is used as a top dressing for the feed, it likewisehelps to ensure uniformity of distribution of the compound across thetop of the dressed feed.

Drinking water and feed effective for increasing lean meat depositionand for improving lean meat to fat ratio are generally prepared bymixing a compound of the present invention with a sufficient amount ofanimal feed to provide from about 10.sub.-3 to about 500 ppm of thecompound in the feed or water.

The preferred medicated swine, cattle, sheep and goat feed generallycontain from about 1 to about 400 grams of a compound of the presentinvention (or combination) per ton of feed, the optimum amount for theseanimals usually being about 50 to about 300 grams per ton of feed.

The preferred poultry and domestic pet feeds usually contain about 1 toabout 400 grams and preferably about 10 to about 400 grams of a compoundof the present invention (or combination) per ton of feed.

For parenteral administration in animals, the compounds of the presentinvention (or combination) may be prepared in the form of a paste or apellet and administered as an implant, usually under the skin of thehead or ear of the animal in which increase in lean meat deposition andimprovement in lean meat to fat ratio is sought.

Paste Formulations may be prepared by dispersing the drug in apharmaceutically acceptable oil such as peanut oil, sesame oil, corn oilor the like.

Pellets containing an effective amount of a compound of the presentinvention, pharmaceutical composition, or combination may be prepared byadmixing a compound of the present invention or combination with adiluent such as carbowax, carnuba wax, and the like, and a lubricant,such as magnesium or calcium stearate, may be added to improve thepelleting process.

It is, of course, recognized that more than one pellet may beadministered to an animal to achieve the desired dose level which willprovide the increase in lean meat deposition and improvement in leanmeat to fat ratio desired. Moreover, implants may also be madeperiodically during the animal treatment period in order to maintain theproper drug level in the animal's body.

The present invention has several advantageous veterinary features. Forthe pet owner or veterinarian who wishes to increase leanness and/ortrim unwanted fat from pet animals, the instant invention provides themeans by which this may be accomplished. For poultry, beef and swinebreeders, utilization of the method of the present invention yieldsleaner animals that command higher sale prices from the meat industry.

EXAMPLES

Unless specified otherwise, starting materials are generally availablefrom commercial sources such as Aldrich Chemicals Co. (Milwaukee, Wis.),Lancaster Synthesis, Inc. (Windham, N.H.), Acros Organics (Fairlawn,N.J.), Maybridge Chemical Company, Ltd. (Cornwall, England) and TygerScientific (Princeton, N.J.). Certain common abbreviations and acronymshave been employed which may include: AcOH (acetic acid), DBU(1,8-diazabicyclo[5.4.0]undec-7-ene), CDI (1,1′-carbonyldiimidazole),DCM (dichloromethane), DEA (diethylamine), DIPEA(N,N-diisopropylethylamine), DMAP (4-dimethylaminopyridine), DMF(N,N′-dimethylformamide), DMSO (dimethylsulfoxide), EDCI(N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide), Et₂O (diethyl ether),EtOAc (ethyl acetate), EtOH (ethanol), HATU(2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate methanaminium), HBTU(O-benzotriazol-1-yl-N,N,N′,N′-tetramethyluronium hexafluoro phosphate),HOBT (1-hydroxybenzotriazole), IPA (isopropyl alcohol), KHMDS (potassiumhexamethyldisilazane), MeOH (methanol), MTBE (tert-butyl methyl ether),NaBH(OAc)₃ (sodium triacetoxyborohydride), NaHMDS (sodiumhexamethyldisilazane), NMP (N-methylpyrrolidone), SEM([2-(Trimethylsilyl)ethoxy]methyl), TEA (triethylamine), TFA(trifluoroacetic acid), THF (tetrahydrofuran), and T₃P (propanephosphonic acid anhydride).

Reactions were performed in air or, when oxygen- or moisture-sensitivereagents or intermediates were employed, under an inert atmosphere(nitrogen or argon). When appropriate, reaction apparatuses were driedunder dynamic vacuum using a heat gun, and anhydrous solvents(Sure-Seal™ products from Aldrich Chemical Company, Milwaukee, Wis. orDriSolv™ products from EMD Chemicals, Gibbstown, N.J.) were employed.Commercial solvents and reagents were used without further purification.When indicated, reactions were heated by microwave irradiation usingBiotage Initiator or Personal Chemistry Emrys Optimizer microwaves.Reaction progress was monitored using thin layer chromatography (TLC),liquid chromatography-mass spectrometry (LCMS), high performance liquidchromatography (HPLC), and/or gas chromatography-mass spectrometry(GCMS) analyses. TLC was performed on pre-coated silica gel plates witha fluorescence indicator (254 nm excitation wavelength) and visualizedunder UV light and/or with I₂, KMnO₄, CoCl₂, phosphomolybdic acid,and/or ceric ammonium molybdate stains. LCMS data were acquired on anAgilent 1100 Series instrument with a Leap Technologies autosampler,Gemini C₁₈ columns, MeCN/water gradients, and either TFA, formic acid,or ammonium hydroxide modifiers. The column eluent was analyzed usingWaters ZQ mass spectrometer scanning in both positive and negative ionmodes from 100 to 1200 Da. Other similar instruments were also used.HPLC data were acquired on an Agilent 1100 Series instrument usingGemini or XBridge C18 columns, MeCN/water gradients, and either TFA orammonium hydroxide modifiers. GCMS data were acquired using a HewlettPackard 6890 oven with an HP 6890 injector, HP-1 column (12 m×0.2mm×0.33 μm), and helium carrier gas. The sample was analyzed on an HP5973 mass selective detector scanning from 50 to 550 Da using electronionization. Purifications were performed by medium performance liquidchromatography (MPLC) using Isco CombiFlash Companion, AnaLogixIntelliFlash 280, Biotage SP1, or Biotage Isolera One instruments andpre-packed Isco RediSep or Biotage Snap silica cartridges. Chiralpurifications were performed by chiral supercritical fluidchromatography (SFC) using Berger or Thar instruments; ChiralPAK-AD,-AS, -IC, Chiralcel-OD, or -OJ columns; and CO₂ mixtures with MeOH,EtOH, iPrOH, or MeCN, alone or modified using TFA or iPrNH₂. UVdetection was used to trigger fraction collection.

Mass spectrometry data are reported from LCMS analyses. Massspectrometry (MS) was performed via atmospheric pressure chemicalionization (APCI), electrospray Ionization (ESI), electron impactionization (EI) or electron scatter (ES) ionization sources. Protonnuclear magnetic spectroscopy (¹H NMR) chemical shifts are given inparts per million downfield from tetramethylsilane and were recorded on300, 400, 500, or 600 MHz Varian spectrometers. Chemical shifts areexpressed in parts per million (ppm, δ) referenced to the deuteratedsolvent residual peaks. The peak shapes are described as follows: s,singlet; d, doublet; t, triplet; q, quartet; quin, quintet; m,multiplet; br s, broad singlet; app, apparent. Analytical SFC data wereacquired on a Berger analytical instrument as described above. Opticalrotation data were acquired on a PerkinElmer model 343 polarimeter usinga 1 dm cell. Silica gel chromatography was performed primarily using amedium pressure Biotage or ISCO systems using columns pre-packaged byvarious commercial vendors including Biotage and ISCO. Microanalyseswere performed by Quantitative Technologies Inc. and were within 0.4% ofthe calculated values.

Unless otherwise noted, chemical reactions were performed at roomtemperature (about 23 degrees Celsius).

The compounds and intermediates described below were named using thenaming convention provided with ChemBioDraw Ultra, Version 12.0(CambridgeSoft Corp., Cambridge, Mass.). The naming convention providedwith ChemBioDraw Ultra, Version 12.0 are well known by those skilled inthe art and it is believed that the naming convention provided withChemBioDraw Ultra, Version 12.0 generally comports with the IUPAC(International Union for Pure and Applied Chemistry) recommendations onNomenclature of Organic Chemistry and the CAS Index rules. Unless notedotherwise, all reactants were obtained commercially without furtherpurifications or were prepared using methods known in the literature.

The terms “concentrated”, “evaporated”, and “concentrated in vacuo”refer to the removal of solvent at reduced pressure on a rotaryevaporator with a bath temperature less than 60° C. The abbreviation“min” and “h” stand for “minutes” and “hours” respectively. The term“TLC” refers to thin layer chromatography, “room temperature or ambienttemperature” means a temperature between 18 to 25° C., “GCMS” refers togas chromatography-mass spectrometry, “LCMS” refers to liquidchromatography-mass spectrometry, “UPLC” refers to ultra performanceliquid chromatography and “HPLC” refers to high pressure liquidchromatography, “SFC” refers to supercritical fluid chromatography.

Hydrogenation may be performed in a Parr Shaker under pressurizedhydrogen gas, or in Thales-nano H-Cube flow hydrogenation apparatus atfull hydrogen and a flow rate between 1-2 mL/min at specifiedtemperature.

HPLC, UPLC, LCMS, GCMS, and SFC retention times were measured using themethods noted in the procedures.

PREPARATION OF INTERMEDIATES AND EXAMPLES General Procedure A:

Add diisopropylethylamine (3.0 equiv) to a solution of Intermediate 1 orIntermediate 2 (1 equiv) in dimethylformamide or tetrahydrofuran (0.15M). HATU (1.0 equiv) and the appropriate amine (2.0 equiv) were addedsequentially to a vial. The reaction mixture was stirred at roomtemperature for 16 h. The reaction was concentrated and purified bypreparative HPLC to afford the specified product unless otherwise noted.

General Procedure B:

Add oxalyl chloride (2.0 eq, 2M in dichloromethane) to a suspension ofIntermediate 1 or Intermediate 2 (1.0 equiv) in dichloromethane (0.1 M).Dimethylformamide (10 uL) was added to the suspension and the reactionmixture was stirred at room temperature for 30 minutes. The suspensionturned to a solution over this time. The appropriate amine (1.0 equiv)and diisopropylethylamine (2.3 equiv) in dichloromethane (0.5 mL) wasadded to the reaction mixture and stirred at room temperature for 16hours. The reaction was diluted with dichloromethane (2 mL) and washedsequentially with 1N sodium hydroxide (100 uL) and brine (100 uL). Theorganic layer was dried over magnesium sulfate, filtered, and thesolvent was evaporated under reduced pressure. The resulting residue waspurified by preparative HPLC to afford the specified product unlessotherwise noted.

Intermediate 1:2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)pyrimidine-5-carboxylicacid

Step 1: 3-Ethoxypyridine

Cesium carbonate (12 mol, 1.5 equiv) and ethyl iodide (9.7 mol, 1.2equiv) were added to a solution of 3-hydroxypyrdine (8.10 mol, 1.0equiv) in acetone (12 L) at 15° C. The reaction mixture was stirred atroom temperature for 24 hours. The reaction mixture was filtered and theorganic layer was concentrated to give crude product. Ethyl acetate (20L) was added and washed with water (3×5 L). The organic layer was driedover sodium sulfate, filtered and concentrated to give 3-ethoxypyridine(620 g, 62%) as an oil. ¹H NMR (400 MHz, CDCl₃) δ 1.44 (t, 3H), 4.07 (q,2H), 7.15-7.23 (m, 2H), 8.20 (dd, 1H), 8.30 (d, 1H).

Step 2: 3-Ethoxypyridine-1-oxide

m-Chloroperoxybenzoic acid (6.5 mol, 1.3 equiv) was added to a solutionof 3-ethoxypyridine (5.0 mol, 1.0 equiv) in dichloromethane (12 L) at10° C. The reaction mixture was stirred at room temperature for 24hours. Sodium thiosulfate (4 kg, in 5 L of water) was added. Thereaction mixture was stirred at 15° C. for 2 hours. Another portion ofsodium thiosulfate (1.5 kg, in 5 L of water) was added. The reactionmixture was stirred at 15° C. for 1 hour. The mixture was extracted withdichloromethane (16×10 L). The combined organic layers were concentratedto give crude product. The crude product was purified by silica gelcolumn chromatography (dichloromethane:methanol; 100:1-10:1) to give thetitle compound (680 g, 97%) as brown oil. This was further purified bytrituration with petroleum ether (4 L) at room temperature for 24 hoursto give 3-ethoxypyridine-1-oxide (580 g, 83%) as yellow solid. ¹H NMR(400 MHz, CDCl₃) δ 1.41 (t, 3H), 4.02 (q, 2H), 6.84 (dd, 1H), 7.12 (dd,1H), 7.85 (d, 1H), 7.91-7.95 (m, 1H).

Step 3: 2-((5-Bromopyridin-3-yl)oxy)-3-ethoxypyridine

This reaction was carried out in five parallel batches.

Diisopropylethylamine (2.69 mol, 3.7 equiv) andbromotripyrrolidinophosphonium hexafluorophosphate (0.93 mol, 1.3 equiv)were added to a stirred solution of 3-ethoxypyridine-1-oxide (0.72 mol,1.0 equiv) and 3-bromo-5-hydroxypyridine (0.72 mol, 1.0 equiv) intetrahydrofuran (2500 mL) at room temperature. The reaction mixture wasstirred at room temperature for 2 days then the separate batches werecombined to a single batch. The resulting suspension was concentrated todryness and dissolved in dichloromethane (25 L). The organic layer waswashed with 1N sodium hydroxide (15 L), water (3×20 L), and brine (20L). The organic layer was dried over sodium sulfate, filtered andconcentrated to give an oil. The crude oil was purified by silica gelcolumn chromatography (petroleum ether:ethyl acetate; 10:1-1:1) to givecrude product as brown solid. This solid was triturated with methyltert-butyl ether: petroleum ether (1:10; 11 L) to afford2-((5-bromopyridin-3-yl)oxy)-3-ethoxypyridine (730 g, 69%) as off yellowsolid. ¹H NMR (400 MHz, CDCl₃) δ 1.49 (t, 3H), 4.16 (q, 2H), 7.04 (dd,1H), 7.25 (dd, 1H), 7.68-7.73 (m, 2H), 8.44 (d, 1H), 8.49 (d, 1H). MS(ES+) 297.1 (M+H).

Step 4: Ethyl2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)pyrimidine-5-carboxylate

A solution of 2-((5-bromopyridin-3-yl)oxy)-3-ethoxypyridine (300 mmol,1.0 equiv) in tetrahydrofuran (1.3 L) was degassed with nitrogen for 30minutes. Turbo Grignard (390 mmol, 1.3 equiv, 1.3 M in tetrahydrofuran)was added at room temperature at a rate to maintain the internaltemperature below 30° C. The reaction mixture was allowed to cool toroom temperature and stirred for 3 hours. The reaction was cooled to 10°C. and zinc chloride (390 mmol, 1.3 equiv, 1.9 M in2-methyltetrahydrofuran) was added at a rate to maintain the temperaturebelow 15° C. The resulting suspension was warmed to room temperatureuntil all the precipitate was dissolved and then cooled back to 10° C.Ethyl 2-chloropyrimidine-5-carboxylate (360 mmol, 1.2 equiv) anddichloro[bis(2-(diphenylphosphino)phenyl)ether]palladium(II) (6.00 mmol,0.02 equiv) were added as solids. The resulting suspension was degassedwith nitrogen for 30 minutes then heated to 50° C. for 16 hours. Thereaction was worked up under aqueous conditions then treatedsequentially with ethylenediaminetetraacetic acid disodium salt,thiosilica, and charcoal to remove metal impurities. The crude compoundwas recrystallized from methanol (450 mL) to yield ethyl2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)pyrimidine-5-carboxylate(77 g, 70%) as a pale, yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 1.44 (t,3H), 1.50 (t, 3H), 4.19 (q, 2H), 4.46 (q, 2H), 7.00-7.04 (m, 1H), 7.25(s, 1H), 7.71 (d, 1H), 8.59 (s, 1H), 8.66 (d, 1H), 9.32 (s, 2H), 9.55(s, 1H).

Step 5:2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)pyrimidine-5-carboxylicacid

Sodium hydroxide (307 mmol, 1.5 equiv, 4M aqueous) and methanol (50 mL)were added to a suspension of2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)pyrimidine-5-carboxylate(205 mmol, 1.0 equiv) in tetrahydrofuran (300 mL). The resultingsolution was stirred at room temperature for 3 hours. The reactionmixture was diluted with water (400 mL) and extracted with 2:1 diethylether:heptanes (2×300 mL). The aqueous layer was acidified to pH of 4with 4M hydrochloric acid. The resulting suspension was stirred at roomtemperature for 1 hour. The solid was filtered, washed with water, anddried to yield2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)pyrimidine-5-carboxylicacid (69 g, 100%) as a pale, yellow solid. ¹H NMR (400 MHz, DMSO-d₆)51.37 (t, 3H), 4.18 (q, 2H), 7.19 (dd, 1H), 7.58 (dd, 1H), 7.70 (dd,1H), 8.35-8.40 (m, 1H), 8.66 (d, 1H), 9.33 (s, 2H), 9.41 (d, 1H), 13.9(br. s, 1H).

Intermediate 2:3-{5-[(3-Ethoxypyridin-2-yl)oxy]pyridin-3-yl}-1,2,4-triazine-6-carboxylicacid

Step 1. 6-Bromo-1,2,4-triazin-3-amine

Water (120 mL) was added to a mixture of 3-amino-1,2,4-triazine (104mmol, 1.0 equiv) in acetonitrile (120 mL) and stirred at roomtemperature until a brown solution was formed. The mixture was cooled to0° C., treated with N-bromosuccinimide (109 mmol, 1.05 equiv) in aportionwise manner and stirred at 0° C. for 20 min. After warming toroom temperature, the mixture was diluted with ethyl acetate (350 mL)and cooled to 0° C. Sodium carbonate (12 g) was added to the mixture andstirred for 10 min. The two layers were separated and the aqueous phasewas extracted with ethyl acetate (200 mL). The combined organic layerswere washed with aqueous sodium bicarbonate, dried over sodium sulfate,filtered and concentrated to give the 6-bromo-1,2,4-triazin-3-amine(10.5 g, 58%) as a yellow solid. ¹H NMR (400 MHz, CD₃OD) 8.32 (s, 1H).

Step 2. Ethyl 3-amino-1,2,4-triazine-6-carboxylate

In two separate batches, palladium acetate (0.87 mmol, 0.05 equiv) wasadded to a solution of 6-bromo-1,2,4-triazin-3-amine (17 mmol, 1.0equiv), triethylamine (35 mmol, 2.0 equiv) and xantphos (1.40 mmol, 0.08equiv) in ethanol (60 mL) was added. The mixture was degassed withcarbon monoxide and stirred at 85° C. under an atmosphere of carbonmonoxide (16 Psi) for 16 h. The cooled reaction mixture was diluted withethyl acetate (60 mL), filtered through a pad of celite andconcentrated. The crude products from both batches were combined andpurified using column chromatography (ethyl acetate/petroleum ether=3:7)to give ethyl 3-amino-1,2,4-triazine-6-carboxylate (2.5 g, 88%) as ayellow solid. ¹H NMR (400 MHz, CDCl₃) δ 1.49 (t, 3H), 4.50 (q, 2H), 8.79(s, 1H).

Step 3. ethyl 3-chloro-1,2,4-triazine-6-carboxylate

tert-Butyl nitrite (4.5 mmol, 1.5 equiv) was added to a solution ofethyl 3-amino-1,2,4-triazine-6-carboxylate (3.0 mmol, 1.0 equiv) andcopper(II) chloride (3.6 mmol, 1.2 equiv) in acetonitrile (15 mL) in adropwise manner. The resulting mixture was heated at 60° C. for 1 h. Thereaction mixture was cooled to room temperature and treated with coldhydrochloric acid (10 mL, 1N). The mixture was extracted with ethylacetate (3×30 mL) and the combined organic layers were dried over sodiumsulfate, filtered and evaporated. The crude product was purified usingcolumn chromatography eluting with ethyl acetate/petroleum ether (5:95to 1:1) to give ethyl 3-chloro-1,2,4-triazine-6-carboxylate (300 mg,54%) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 1.50 (t, 3H), 4.58 (q,2H), 9.11 (s, 1H).

Step 4. 3-Ethoxy-2-{[5-(tributylstannanyl)pyridin-3-yl]oxy}pyridine

Tetrakis(triphenylphosphine)palladium(0) (0.68 mmol, 0.10 equiv) wasadded to a solution of 2-[(5-bromopyridin-3-yl)oxy]-3-ethoxypyridine(6.8 mmol, 1.0 equiv) and hexabutyldistannane (7.5 mmol, 1.1 equiv) indioxane (40 mL) under an atmosphere of nitrogen. The reaction was heatedto 110° C. and stirred at this temperature for 16 h. The mixture wasquenched with aqueous potassium fluoride and stirred for 1 h. Theresulting suspension was filtered through a pad of celite and thefiltrate was extracted with ethyl acetate (3×60 mL). The combinedorganics were washed with brine, dried over sodium sulfate, filtered andconcentrated. The crude product was purified using column chromatography(ethyl acetate/petroleum ether=0:100 to 1:4) to give3-ethoxy-2-{[5-(tributylstannanyl)pyridin-3-yl]oxy}pyridine (1.6 g, 47%)as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ 0.88 (t, 9H), 1.06-1.11 (m,6H), 1.32 (s, 6H), 1.47-1.58 (m, 9H), 4.17 (q, 2H), 6.99 (dd, 1H), 7.22(dd, 1H), 7.57 (dd, 1H), 7.70 (dd, 1H), 8.37-8.40 (m, 2H).

Step 5. Ethyl3-{5-[(3-ethoxypyridin-2-yl)oxy]pyridin-3-yl}-1,2,4-triazine-6-carboxylate

Tetrakis(triphenylphosphine)palladium(0) (0.05 mmol, 0.05 equiv) wasadded to a mixture of3-ethoxy-2-{[5-(tributylstannanyl)pyridin-3-yl]oxy}pyridine (0.99 mmol,1.0 equiv) and ethyl 3-chloro-1,2,4-triazine-6-carboxylate (0.99 mmol,1.0 equiv) in dioxane (8 mL). The vial was degassed to remove oxygen bybubbling through nitrogen gas gently for 2 min. The vial was thenstirred at 115° C. for 30 min under microwave irradiation. The reactionmixture was cooled to room temperature, treated with aqueous potassiumfluoride, and stirred for 1 h. The suspension was filtered through a padof celite and the filtrate was extracted with ethyl acetate (3×30 mL).The combined organic layers were washed with brine, dried over sodiumsulfate and concentrated. The crude material was purified using columnchromatography (ethyl acetate/petroleum ether=1:4 to 1:1) to give ethyl3-{5-[(3-ethoxypyridin-2-yl)oxy]pyridin-3-yl}-1,2,4-triazine-6-carboxylate(150 mg, 41%) as a yellow oil. MS (ES+) 368.0 (M+H).

Step 6.3-{5-[(3-ethoxypyridin-2-yl)oxy]pyridin-3-yl}-1,2,4-triazine-6-carboxylicacid

Sodium hydroxide (1.0 mmol, 20 equiv, 2M) was added to a solution ofethyl3-{5-[(3-ethoxypyridin-2-yl)oxy]pyridin-3-yl}-1,2,4-triazine-6-carboxylate(0.053 mmol, 1.0 equiv) in methanol (1 mL) at room temperature. Thesolution was stirred for 1 h. The mixture was concentrated to removemethanol, diluted with water, and extracted with dichloromethane (2×15mL). The aqueous layer was acidified to pH=5 with 2 N hydrochloric acidand extracted with ethyl acetate (3×10 mL). The combined organic layerswere washed with brine (20 mL), dried over anhydrous sodium sulfate,filtered and concentrated to dryness to give the product (15 mg, 83%) asa light yellow solid. MS (ES+) 339.9 (M+H).

Intermediate 3:2-{5-[(3-ethoxypyrazin-2-yl)oxy]pyridin-3-yl}pyrimidine-5-carboxylicacid

Step 1. 2-[(5-bromopyridin-3-yl)oxy]-3-ethoxypyrazine

5-Bromopyridin-3-ol (32 mmol, 1.0 equiv) and cesium carbonate (39 mmol,1.3 equiv) were added to a solution of 2-chloro-3-ethoxypyrazine (32mmol, 1.0 equiv) in N-methyl-2-pyrrolidone (250 mL). The reactionmixture was stirred at 150° C. for 1 h. The cooled reaction mixture waspoured into water (300 mL) and extracted with ethyl acetate (3×250 mL).The combined organic layers were dried over sodium sulfate, filtered andconcentrated. The crude product was purified using column chromatographyeluting with petroleum ether/ethyl acetate (0% to 100%) to give thetitle compound (5.0 g, 54%) as a white solid. ¹H NMR (400 MHz, CDCl₃) δ1.50 (t, 3H), 4.54 (q, 2H), 7.59 (d, 1H), 7.77 (t, 1H), 7.83 (d, 1H),8.49 (d, 1H), 8.56 (d, 1H).

Step 2. Ethyl2-{5-[(3-ethoxypyrazin-2-yl)oxy]pyridin-3-yl}pyrimidine-5-carboxylate

[1,1′-Bis(diphenylphosphino) ferrocene] dichloropalladium (II) complexwith dichloromethane (124 mg, 0.05 eq) was added to a suspension of2-((5-bromopyridin-3-yl)oxy)-3-ethoxypyrazine (3.4 mmol, 1.0 equiv),bis(pinacolato)diboron (4.1 mmol, 1.2 eq), and potassium acetate (13mmol, 4.0 eq) in dioxane (5 mL). The reaction mixture was purged withnitrogen and stirred at 100° C. for 2 hours. The reaction was cooled toroom temperature and quenched with water (50 mL). The mixture wasextracted with ethyl acetate (25 mL) and washed with brine (3×50 mL).The organic layer was concentrated to provide2-ethoxy-3-((5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl)oxy)pyrazine(1.4 g, 110%) as a black oil that was used directly in the next step.Ethyl-2-chloropyrimidine-5-carboxylate (930 mg, 1.2 eq), potassiumcarbonate (1.1 g, 2.0 equiv), and [1,1′-bis(diphenylphosphino)ferrocene] dichloropalladium (II) complex with dichloromethane (150 mg,0.05 eq) were added to a solution of2-ethoxy-3-((5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl)oxy)pyrazine(1.4 g, 4.1 mmol) in dioxane (5 mL). The black suspension was flushedwith nitrogen and stirred at 80° C. for 16 hours. The reaction wascooled to room temperature and quenched with water (50 mL). The mixturewas extracted with ethyl acetate (25 mL), washed with brine (3×50 mL)and dried to black residue. The crude was purified by flashchromatography (ethyl acetate in petroleum ether) to afford ethyl2-{5-[(3-ethoxypyrazin-2-yl)oxy]pyridin-3-yl}pyrimidine-5-carboxylate(700 mg, 46%) as a yellow solid. MS (ES+) 367.9 (M+H).

Step 4.2-{5-[(3-ethoxypyrazin-2-yl)oxy]pyridin-3-yl}pyrimidine-5-carboxylicacid

Sodium hydroxide (6.8 mmol, 5.0 equiv, 2M) was added to ethyl2-{5-[(3-ethoxypyrazin-2-yl)oxy]pyridin-3-yl}pyrimidine-5-carboxylate(1.4 mmol, 1.0 equiv) in ethanol (5 mL). The reaction was stirred at 30°C. for 16 hours. Methyl-tert-butyl ether (30 mL) was added to thereaction mixture and the resulting solid was filtered and dried toprovide2-{5-[(3-ethoxypyrazin-2-yl)oxy]pyridin-3-yl}pyrimidine-5-carboxylicacid (500 mg, 101%) as the sodium salt. ¹H NMR (300 MHz, DMSO-d₆) δ 1.42(t, 3H), 4.48 (q, 2H), 7.67 (s, 1H), 7.93 (s, 1H), 8.54 (s, 1H), 8.74(s, 1H), 9.33 (s, 2H), 9.45 (s, 1H).

Intermediate 4:2-(5-((3-(2-Fluoroethoxy)pyridin-2-yl)oxy)pyridin-3-yl)pyrimidine-5-carboxylicacid

Step 1: 3-(2-Fluoroethoxy)pyridine

Potassium carbonate (1.5 g, 2.0 equiv) was added to a solution of3-hydroxypyridine (500 mg, 1 equiv) and 1-fluoro-2-iodoethane (920 mg,1.0 equiv) in dimethylformamide (10 mL). The suspension was stirred at30° C. for 16 hours. The reaction was diluted with 10% methanol indichloromethane (90 mL) and washed with water (20 mL). The organic layerwas washed with brine, dried over sodium sulfate and concentrated. Thecrude material was purified by flash chromatography (gradient: 0-4.5%methanol in dichloromethane) to afford 3-(2-fluoroethoxy)pyridine (500mg, 67%) as a brown oil. ¹H NMR (400 MHz, CDCl₃) b 4.19-4.33 (m, 2H),4.67-4.74 (m, 1H), 4.79-4.86 (m, 1H), 7.22 (dd, 2H), 8.24 (t, 1H), 8.34(t, 1H).

Step 2: 3-(2-Fluoroethoxy)pyridine 1-oxide

m-Chloroperoxybenzoic acid (2.49 g, 1.2 equiv) was added to a solutionof 3-(2-fluoroethoxy)pyridine (1.7 g, 1.0 equiv) in dichloromethane (30mL). The reaction was stirred at room temperature for 16 hours. Thereaction was purified directly by flash chromatography (gradient: 0-5%methanol in dichloromethane) to yield 3-(2-fluoroethoxy)pyridine 1-oxide(1.2 g, 63%) as a white solid. ¹H NMR (400 MHz, CDCl₃) δ 4.16-4.22 (m,1H), 4.23-4.29 (m, 1H), 4.66-4.73 (m, 1H), 4.77-4.84 (m, 1H), 6.90 (dd,1H), 7.17 (dd, 1H), 7.87-7.94 (m, 1H), 7.99 (t, 1H).

Step 3: 2-((5-Bromopyridin-3-yl)oxy)-3-(2-fluoroethoxy)pyridine

Diisopropylethylamine (5.2 mL, 3.8 equiv) was added to a solution of3-(2-fluoroethoxy)pyridine 1-oxide (1.2 g, 1.0 equiv),3-bromo-5-hydroxypyridine (1.3 g, 1.0 equiv) andbromotripyrrolidinophosphonium hexafluorophosphate (4.6 g, 1.3 equiv) intetrahydrofuran (25 mL) at 13° C. The mixture was stirred at roomtemperature for 16 hours. The reaction was quenched with water (20 mL)and extracted with ethyl acetate (50 mL). The combined organics werewashed with saturated ammonium chloride (3×20 mL) and brine (100 mL),dried over sodium sulfate, and concentrated. The crude material waspurified by flash chromatography (gradient: 0-70% ethyl acetate inpetroleum ether) to provide2-((5-Bromopyridin-3-yl)oxy)-3-(2-fluoroethoxy)pyridine (1.9 g, 81%) asa yellow oil. ¹H NMR (400 MHz, CDCl₃) δ 4.28-4.33 (m, 1H), 4.34-4.41 (m,2H), 4.70-4.78 (m, 1H), 4.82-4.82 (m, 1H), 7.05 (dd, 1H), 7.31 (dt, 1H),7.69-7.73 (m, 1H), 7.74-7.80 (m, 2H), 8.44 (dd, 1H), 8.48-8.52 (m, 2H).

Step 4:2-(5-((3-(2-Fluoroethoxy)pyridin-2-yl)oxy)pyridin-3-yl)pyrimidine-5-carboxylicacid

Bis(pinacolato)diboron (580 mg, 1.2 equiv), potassium acetate (560 mg,3.0 equiv), and [1,1′-Bis(diphenylphosphino) ferrocene]dichloropalladium (11) complex with dichloromethane (70 mg, 0.05 equiv)was added to a solution of2-((5-Bromopyridin-3-yl)oxy)-3-(2-fluoroethoxy)pyridine (600 mg, 1.0equiv) in dioxane (10 mL) at room temperature. The reaction was stirredat 100° C. for 16 hours. The reaction mixture was diluted with ethylacetate (60 mL), washed with water (20 mL) and brine, then concentratedto a residue. The residue was diluted with dioxane (15 mL) and water (5mL). Ethyl 2-chloropyrimidine-5-carboxylate (310 mg, 1.0 equiv),[1,1′-Bis(diphenylphosphino) ferrocene]dichloropalladium (II) complexwith dichloromethane (61 mg, 0.05 equiv), and potassium carbonate (460mg, 2.0 equiv) were added to the reaction mixture and the resultingsuspension was stirred at 80° C. for 16 hours. The suspension wasfiltered and then partitioned between ethyl acetate (30 mL) and water(50 mL). The aqueous layer was extracted with ethyl acetate (2×30 mL).The combined organic layers were washed with brine (50 mL), dried oversodium sulfate, filtered, and concentrated to provide crude materialthat was purified by prep-TLC (5% methanol in dichloromethane) to afford2-(5-((3-(2-fluoroethoxy)pyridin-2-yl)oxy)pyridin-3-yl)pyrimidine-5-carboxylicacid (80 mg, 13%). MS (ES+) 357.0 (M+H).

Example 1:(S)-2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(tetrahydrofuran-3-yl)pyrimidine-5-carboxamide

Oxalyl chloride (13.8 mL, 160 mmol, 1.2 equiv) and dimethylformamide(0.510 mL, 6.65 mmol, 0.05 equiv) were added to a suspension of2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)pyrimidine-5-carboxylicacid (45.0 g, 133 mmol, 1.0 equiv) in dichloromethane (500 mL). Thesuspension was stirred for 2 hours when a solution was achieved. Thereaction mixture was concentrated to yield crude acid chloride as a redsolid. A solution of (S)-tetrahydrofuran-3-amine (12.2 g, 140 mmol, 1.05equiv) and diisopropylethylamine (51.0 mL, 293 mmol, 2.2 equiv) intetrahydrofuran (100 mL) was added dropwise to a solution of the crudeacid chloride in dichloromethane (200 mL) at 0° C. The reaction wasallowed to warm to room temperature and stirred for 16 hours. Water (1.0L) and ethyl acetate (600 mL) were added and the organic layer wasseparated, washed with saturated sodium bicarbonate, dried overmagnesium sulfate, and filtered. The filtrate was treated with activatedcharcoal (20 g) was stirred at 65° C. for 20 minutes. The suspension wasfiltered warm and filtrate was concentrated to a pale, yellow solidwhich was recrystallized from methanol in ethyl acetate (1:4, 1 L) toyield(S)-2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(tetrahydrofuran-3-yl)pyrimidine-5-carboxamide(43.5 g, 81%) as a colorless solid. The title compound was combined withprevious batches (108.7 g, 266.8 mmol) prepared in the same manner andslurried with ethyl acetate (1.0 L) at 80° C. for 4 hours. Thesuspension was allowed to cool to room temperature and stirred for 4days. The solid was filtered, washed with ethyl acetate (3×200 mL) anddried under high vacuum at 50° C. for 24 hours to yield(S)-2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(tetrahydrofuran-3-yl)pyrimidine-5-carboxamide(100.5 g, 92%) as a colorless solid. ¹H NMR (300 MHz, DMSO-d₆) δ 1.38(t, 3H), 1.89-1.98 (m, 1H), 2.15-2.26 (m, 1H), 3.65 (dd, 1H), 3.70-3.78(m, 1H), 3.85-3.92 (m, 2H), 4.18 (q, 2H), 4.46-4.55 (m, 1H), 7.18 (dd,1H), 7.58 (dd, 1H), 7.69 (dd, 1H), 8.37 (dd, 1H), 8.64 (d, 1H), 8.95 (d,1H), 9.28 (s, 2H), 9.39 (d, 1H). MS (ES+) 408.4 (M+H). Melting point177.5° C. Elemental analysis for C₂₁H₂₁N₅O₄: calculated C, 61.91; H,5.20; N, 17.19; found C, 61.86; H, 5.18; N, 17.30.

The solid form from this procedure was characterized by Powder X-raydiffraction (PXRD) analysis and assigned as Form 1.

Alternative preparation for(S)-2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(tetrahydrofuran-3-yl)pyrimidine-5-carboxamide(Example 1)

A 100 mL reactor was charged with acetonitrile (35 mL),2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)pyrimidine-5-carboxylicacid (5.0 g, 15 mmol) and (S)-tetrahydrofuran-3-amine hydrochloride (2.2g, 18 mmol, 1.2 equiv). Diisopropylethylamine (18 mL, 103 mmol, 7.0equiv) was charged while maintaining the temperature at 20° C. to 30° C.A solution of propane phosphonic acid anhydride (T3P) in acetonitrile(21 mL, 30 mmol, 2.0 equiv) was charged at a rate that maintained thetemperature below 45° C. The reactor was heated to 40±5° C. for 1 hourthen sampled for reaction completion. The reaction was cooled to 20° C.to 25° C. and tetrahydrofuran (25 mL) was added. A solution of sodiumbicarbonate (0.5M, 40 mL) was charged and the mixture was stirred for 1hour. The pH was checked and measured at 8.5. Ethyl acetate (40 mL) wasadded and the mixture stirred for 15 minutes. The mixture was settledand the phases split. The aqueous layer was transferred to a separatoryfunnel and back extracted with ethyl acetate (100 mL). The organicphases were combined and washed with water (40 mL). The organic layerwas transferred to a 100 mL reactor in portions and concentrated undervacuum to a low volume. Methyl ethyl ketone (100 mL) was added and themixture was concentrated to a final volume of approximately 60 mL.Vacuum was removed and the slurry was heated to reflux and held untilthe solids were washed down the reactor walls. The slurry was cooled to15° C. over 2 hours and granulated overnight. The solids were isolatedby filtration, washing the reactor and cake twice with methyl ethylketone (10 mL each). The solids were dried in a vacuum oven at 50° C. toyield 4.86 g (81%) of the desired product. The solid form from thisprocedure was characterized by PXRD analysis and assigned as Form 2.

Conversion of the Form 2 to the Form 1

To a 100 mL reactor was charged Form 2 of(S)-2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(tetrahydrofuran-3-yl)pyrimidine-5-carboxamide(Example 1) (10.0 g, 24.6 mmol, 1.00 equiv.), Methyl ethyl ketone (8.8mL/g), 88.0 mL) and water (1.2 mL/g, 12.0 mL). The reactor was heated to50° C. over 30 minutes. A complete solution appeared at approximately44° C. The reactor was cooled to 40° C. over 30 minutes then seed Form 1of(S)-2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(tetrahydrofuran-3-yl)pyrimidine-5-carboxamide(Example 1) (0.050 g, 0.123 mmol, 0.0050 equiv.) was charged. Afterseeding, the hazy slurry was stirred for 1 hour before cooling to 5° C.over 2 hours and then stirred at 5° C. for 12 hours. An in processcontrol sample was pulled and characterized by PXRD analysis to confirmthe solids were Form 1. The slurry was filtered, and the reactor andcake was washed with 0° C. methyl ethyl ketone (2.5 mL/g, 25 mL). Thesolids were dried in a vacuum oven at 50° C. to yield 8.15 g (81.5%) ofthe desired product. PXRD patterns of the desired product wereconsistent with Form 1.

Powder X-Ray Diffraction:

Powder X-ray diffraction analysis was conducted using a Bruker AXS D8Advance diffractometer equipped with a Cu radiation source (Kα-averagewavelength of 1. 54056 Å), equipped with a twin primary utilizing agobel mirror. Diffracted radiation was detected by a PSD-Lynx Eyedetector. Both primary and secondary equipped with 2.5 soller slits. TheX-ray tube voltage and amperage were set at 40 kV and 40 mArespectively. Data was collected in the Theta-Theta goniometer in alocked couple scan from 3.0 to 40.0 degrees 2-Theta with 1000 stepsusing a scan speed of 6 seconds per step. Samples were prepared byplacement in a silicon low background sample holder (C79298A3244B261).Data were collected using Bruker DIFFRAC Plus software. Analysisperformed by EVA diffract plus software.

The PXRD data file was not processed prior to peak searching. Using thepeak search algorithm in the EVA software, peaks were selected with athreshold value of 5 and a width value of 0.2. The output of automatedassignments was visually checked to ensure validity and adjustmentsmanually made if necessary. Peaks with relative intensity of ≥3% weregenerally chosen. The peaks which were not resolved or were consistentwith noise were also discarded. A typical error associated with the peakposition from PXRD stated in USP is within +/−0.2° (USP-941).

TABLE 1 Key PXRD peaks to characterize crystalline material of Example 1Form 1 of Example 1 Form 2 of Example 1 Angle 2Θ (°) Angle 2Θ (°) 5.3,7.7, 15.4 6.5, 9.3, 13.6

FIG. 1 is a characteristic x-ray powder diffraction pattern showingcrystalline form 1 of Example 1 (Vertical Axis: Intensity (CPS);Horizontal Axis: Two theta (degrees)).

FIG. 2 is a characteristic x-ray powder diffraction pattern showingcrystalline form 2 of Example 1 (Vertical Axis: Intensity (CPS);Horizontal Axis: Two theta (degrees)).

Example 2:(R)-2-(5-((3-Ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(tetrahydrofuran-3-yl)pyrimidine-5-carboxamide

The title compound was prepared using general method A with Intermediate1 (0.31 mmol, 1.0 equiv) and (R)-(+)-tetrahydro-3-furylaminetoluenesulfonate salt (124 mg, 1.5 eq). The crude product was purifiedby flash chromatography using ethyl acetate in heptanes to yield(R)-2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(tetrahydrofuran-3-yl)pyrimidine-5-carboxamide(91 mg, 70%). ¹H NMR (400 MHz, DMSO-d₆) δ 1.38 (t, 3H), 1.89-1.98 (m,1H), 2.15-2.28 (m, 1H), 3.5 (dd, 1H), 3.70-3.78 (m, 1H), 3.85-3.92 (m,2H), 4.19 (q, 2H), 4.46-4.55 (m, 1H), 7.19 (dd, 1H), 7.58 (dd, 1H), 7.69(dd, 1H), 8.37 (dd, 1H), 8.64 (d, 1H), 8.96 (d, 1H), 9.28 (s, 2H), 9.39(d, 1H). MS (ES+) 408.3 (M+H).

Examples 3.1-3.7

The Examples in Table 2 were prepared by the general procedure A usingthe appropriate starting materials and analyzed by the methods describedbelow. The R³ variable, intermediates (which varies R¹, D¹, and/or D²)used, and analysis method is noted in Table 2.

Analytical Methods:

Method A: Xbridge C18, 2.1×50 mm, 5 μm, 40° C., Mobile Phase A 0.0375%TFA in water, Mobile Phase B 0.01875% TFA in acetonitrile, Gradient:0.00 min 1% B, 0.60 min 5% B, 4.00 min 100% B, 0.8 mL/min, API-ES+.

Method B: Xbridge C18, 2.1×50 mm, 5 μm, 40° C., Mobile Phase A 0.05%NH₄OH in water, Mobile phase B 100% acetonitrile, Gradient 0.00 min 5%B, 3.40 min 100% B, 0.8 mL/min, API-ES+.

Method C: Waters Atlantis dC18 4.6×50, 5 um, Mobile phase A: 0.05% TFAin water (v/v); Mobile phase B: 0.05% TFA in acetonitrile (v/v),Gradient: 95.0% H₂O/5.0% Acetonitrile linear to 5% H₂O/95% Acetonitrilein 4.0 min, HOLD at 5% H₂O/95% Acetonitrile to 5.0 min. Flow: 2 mL/min

Method D: Waters XBridge C18 4.6×50, 5 um, Mobile phase A: 0.03% NH₄OHin water (v/v); Mobile phase B: 0.03% NH₄OH in acetonitrile (v/v),Gradient: 95.0% H₂O/5.0% Acetonitrile linear to 5% H₂O/95% Acetonitrilein 4.0 min, HOLD at 5% H₂O/95% Acetonitrile 15 to 5.0 min. Flow: 2mL/min.

Method E: Xtimate C18 5×30 mm, 3 um

Mobile phase A: 0.1% TFA in water, Mobile Phase B: acetonitrile,Gradient: 0.00 min 1% B, 1 min 5% B, 5 min 100% B, 8 min 1% B. Flowrate: 1.2 mL/min

Method F: LCMS E(4-302) XBridge C18 2.1*50 mm, 5 um

Mobile phase: 1.0% acetonitrile in water (0.1% formic acid) to 5%acetonitrile in water (0.1% formic acid) in 0.6 min; then from 5.0%acetonitrile

in water (0.1% formic acid) to 100% acetonitrile (0.1% formic acid) in3.4 minutes; then back to 1.0% acetonitrile in water (0.1% formic acid)till 4.3 min, and hold 0.7 minutes. Flow rate: 0.8 ml/min

Method G: Xbridge C18, 2.0×50 mm, 5 μm, 40° C., Mobile Phase A 10 mMNH₄HCO₃ in water, Mobile phase B 100% acetonitrile, Gradient 1.0% B to5% B in 0.6 min, 100% B in 3.4 minutes; then back to 1.0% B within 0.3min. Flow rate: 0.8 ml/min

TABLE 2 MS Retention Time Compound Intermediate (ES+) (min) and ExampleName R³ Number (M + H) analytical method 3.1 2-(5-((3- ethoxypyridin- 2-yl)oxy)pyridin- 3-yl)-N-(3- methyl-1,1-

1 470 2.310 Method C dioxidotetra- hydrothiophen-3- yl)pyrimidine-5-carboxamide 3.2 N-(1,3- dihydroxy-2- methylpropan- 2-yl)-2-(5-((3-ethoxypyridin-

1 426 1.683 Method D 2- yl)oxy)pyridin- 3-yl)pyrimidine- 5-carboxamide3.3 N-(1,1- dioxidotetra- hydrothiophen-3- yl)-2-(5-((3- ethoxypyridin-2-

1 456 3.319 Method E yl)oxy)pyridin- 3-yl)pyrimidine- 5-carboxamide 3.4(S)-3-(5-((3- ethoxypyridin- 2- yl)oxy)pyridin- 3-yl)-N- (tetrahydro-

2 409 2.517 Method F furan-3-yl)-1,2,4- triazine-6- carboxamide 3.52-(5-((3- ethoxypyrazin- 2- yl)oxy)pyridin- 3-yl)-N-(1-

3 411 2.274 Method C hydroxy-2- methylpropan- 2-yl)pyrimidine-5-carboxamide 3.6 2-(5-((3- ethoxypyridin- 2- yl)oxy)pyridin-3-yl)-N-(2-

1 472 2.548 Method F methyl-1- (methylsulfonyl) propan-2-yl)pyrimidine-5- carboxamide 3.7 (S)-2-(5-((3-(2- fluoroethoxy)pyridin-2- yl)oxy)pyridin- 3-yl)-N- (tetrahydro-

4   425.9 2.325 Method F furan-3- yl)pyrimidine-5- carboxamide

Example 4:N-(2-cyanopropan-2-yl)-2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)pyrimidine-5-carboxamide

The title compound was prepared following general procedure B withIntermediate 1 (1.0 g, 1.0 equiv) and 2-amino-2-methylpropionitrile (436mg, 1.1 eq). The crude product was purified by flash chromatography(50-100% ethyl acetate in heptanes) and treated with charcoal. Theresulting residue was recrystallized from ethyl acetate (8 mL) to yieldN-(2-cyanopropan-2-yl)-2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)pyrimidine-5-carboxamide(1.1 g, 92%) as a colorless solid. ¹H NMR (400 MHz, DMSO-d₆) δ 1.38 (t,3H), 1.74 (s, 6H), 4.18 (q, 2H), 7.18 (dd, 1H), 7.57 (dd, 1H), 7.69 (dd,1H), 8.38 (dd, 1H), 8.66 (d, 1H), 9.18 (br. s, 1H), 9.30 (s, 2H), 9.40(d, 1H). MS (ES+) 405.3 (M+H).

Example 5:2-(5-((3-Ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(1-hydroxy-2-methylpropan-2-yl)pyrimidine-5-carboxamide

The title compound was prepared following general procedure A withIntermediate 1 (260 mg, 1.0 equiv) and 2-amino-2-methylpropan-1-ol (103mg, 1.5 eq). The crude product was purified by flash chromatography(50-100% ethyl acetate in heptanes) and recrystallized from ethylacetate: methanol to yield2-(5-((3-Ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(1-hydroxy-2-methylpropan-2-yl)pyrimidine-5-carboxamide(236 mg, 75%) as a colorless solid. ¹H NMR (400 MHz, DMSO-d₆) δ 1.33 (s,6H), 1.38 (t, 3H), 3.55 (d, 2H), 4.18 (q, 2H), 4.83 (t, 1H), 7.18 (dd,Hz, 1H), 7.58 (d, 1H), 7.70 (d, 1H), 8.04 (br. s, 1H), 8.34-8.37 (m,1H), 8.64 (d, 1H), 9.22 (s, 2H), 9.39 (d, 1H). MS (ES+) 410.3 (M+H).

Table 3 includes the hepatic clearance profile (CI_(int, app) in HLM) ofExample 5, which showed significant improvement when compared withExample 19.21 in WO2015140658. This significantly reduced clearance ismuch greater than would have been predicted given the difference in thestructures and DGAT2 IC₅₀ values of Example 5 and Example 19.21 ofWO2015140658.

TABLE 3 DGAT2 Potency and Metabolic Clearance in HLM CI_(int, app) DGAT2in HLM Example No. Structure IC₅₀ [nM] [μL/min/mg] Example 5

14 18 W02015140658 Example 19.21

  7.9 121 

Examples 6a and 6b:(R)-2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(3-(hydroxymethyl)tetrahydrofuran-3-yl)pyrimidine-5-carboxamideand(S)-2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(3-(hydroxymethyl)tetrahydrofuran-3-yl)pyrimidine-5-carboxamide

The title compounds were prepared using general procedure A withIntermediate 1 (840 mg, 1 equiv) and(3-aminotetrahydrofuran-3-yl)methanol (320 mg, 1.1 eq). The crudeproduct was passed through a plug of silica to yield the racemate as ayellow solid which was separated by chiral SFC purification: Chiral TechAD-H 250 mm×4.6 mm 5 u; Isocratic 70% A: carbon dioxide; Mobile phase in30% B: 0.2% isopropylamine in isopropanol (v/v). Flow: 60 mL/min; backpressure=120 Bar. The first peak to elute off the column is Example 6a,and the second peak is Example 6b.

Enantiomer 1:

SFC retention time=9.31 minutes. (330 mg) This compound was furtherpurified by column chromatography (methanol in dichloromethane) thenrecrystallized from methanol in ethyl acetate to afford enantiomer 6a(218 mg, 20%) as a colorless solid. ¹H NMR (400 MHz, CD₃OD) δ 1.40 (t,3H), 2.23 (dt, 1H), 2.29-2.37 (m, 1H), 3.82-3.91 (m, 2H), 3.92-3.95 (m,2H), 3.96-4.04 (m, 2H), 4.17 (q, 2H), 7.19 (dd, 1H), 7.52 (dd, 1H), 7.71(dd, 1H), 8.49 (dd, 1H), 8.52 (d, 1H), 9.22 (s, 2H), 9.41 (d, 1H). MS(ES+) 438.3 (M+H).

Enantiomer 2:

SFC retention time=9.59 minutes (300 mg). This compound was furtherpurified by column chromatography (methanol in dichloromethane) thenrecrystallized from methanol in ethyl acetate to afford enantiomer 6b(205 mg, 19%) as a colorless solid. ¹H NMR (400 MHz, CD₃OD) δ 1.40 (t,3H), 2.23 (dt, 1H), 2.29-2.37 (m, 1H), 3.82-3.91 (m, 2H), 3.92-3.95 (m,2H), 3.96-4.04 (m, 2H), 4.17 (q, 2H), 7.19 (dd, 1H), 7.52 (dd, 1H), 7.71(dd, 1H), 8.49 (dd, 1H), 8.52 (d, 1H), 9.22 (s, 2H), 9.41 (d, 1H). MS(ES+) 438.3 (M+H).

Example 7:3-{5-[(3-ethoxypyridin-2-yl)oxy]pyridin-3-yl}-N-(1-hydroxy-2-methylpropan-2-yl)-1,2,4-triazine-6-carboxamide

The title compound was prepared according to general procedure A usingIntermediate 2 (15 mg, 0.04 mmol) and 2-amino-2-methyl-1-propanol (4.73mg, 0.053 mmol). The crude product was purified by reverse-phasepreparatory HPLC to give the title compound (2.8 mg, 16%) as a lightyellow solid. ¹H NMR (400 MHz, CD₃OD) δ 1.40 (t, 3H), 1.47 (s, 6H), 3.70(s, 2H), 4.16 (q, 2H), 7.20 (dd, 1H), 7.53 (dd, 1H), 7.71 (dd, 1H),8.56-8.58 (m, 1H), 8.61 (d, 1H), 9.32 (s, 1H), 9.48 (d, 1H). MS (ES+)411.0 (M+H).

Example 8:(S)-2-(5-((3-Ethoxy-5-fluoropyridin-2-yl)oxy)pyridin-3-yl)-N-(tetrahydrofuran-3-yl)pyrimidine-5-carboxamide

Step 1: 3-Ethoxy-5-fluoropyridine-1-oxide

Cesium carbonate (21.6 g, 3.0 equiv) was added to a solution of5-fluoro-3-pyridinol (25 g, 1.0 equiv) and ethyl iodide (3.8 g, 1.1equiv). The reaction was stirred at room temperature for 16 hours. Themixture was filtered and concentrated to afford3-ethoxy-5-fluoropyridine (3.1 g, 100%) as a yellow oil which was usedwithout further purification. m-Chloroperoxybenzoic acid (5.7 g, 1.5equiv) was added to a solution of 3-ethoxy-5-fluoropyridine (3.1 g, 1.0equiv) in dichloromethane (50 mL). The reaction was stirred at roomtemperature for 16 hours. The reaction was purified directly by flashchromatography (gradient: 0-5% methanol in dichloromethane) to yield3-ethoxy-5-fluoropyridine-1-oxide (3.30 g, 95%) as a white solid. ¹H NMR(400 MHz, CD₃OD) δ 1.40 (t, 3H), 4.14 (q, 2H), 7.20 (dt, 1H), 7.96-7.98(m, 1H), 8.04-8.08 (m, 1H).

Step 2: 2-((5-Bromopyridin-3-yl)oxy)-3-ethoxy-5-fluoropyridine

Diisopropylethylamine (3.08 g, 3.8 equiv) was added to a solution of3-ethoxy-5-fluoropyridine-1-oxide (1.0 g, 1.0 equiv) andbromotripyrrolidinophosphonium hexafluorophosphate (3.86 g, 1.3 equiv)in tetrahydrofuran (60 mL) at 0° C. The mixture was stirred at roomtemperature for 16 hours. The reaction was quenched with water (150 mL)and extracted with ethyl acetate (3×50 mL). The combined organics werewashed with brine (100 mL), dried over sodium sulfate, and concentrated.The crude material was purified by flash chromatography (gradient: 4-24%ethyl acetate in petroleum ether) to provide2-((5-bromopyridin-3-yl)oxy)-3-ethoxy-5-fluoropyridine (200 mg, 10%) asa white solid. ¹H NMR (400 MHz, CDCl3) δ 1.48 (t, 3H), 4.12 (q, 2H),7.04 (dd, 1H), 7.56-7.59 (m, 1H), 7.66-7.67 (m, 1H), 8.41-8.42 (m, 1H),8.48-8.50 (m, 1H).

Step 3: Ethyl2-(5-((3-ethoxy-5-fluoropyridin-2-yl)oxy)pyridin-3-yl)pyrimidine-5-carboxylate

Bis(pinacolato)diboron (243 mg, 1.2 equiv), potassium acetate (235 mg,3.0 equiv), and [1,1′-Bis(diphenylphosphino) ferrocene]dichloropalladium (11) complex with dichloromethane (29 mg, 0.05 equiv)was added to a solution of2-((5-bromopyridin-3-yl)oxy)-3-ethoxy-5-fluoropyridine (250 mg, 1.0equiv) in dioxane (5 mL) at room temperature. The reaction was stirredat 100° C. for 2 hours and then cooled to room temperature. The reactionmixture was diluted with ethyl acetate (60 mL) and filtered throughcelite. The filtrate was concentrated to a residue then diluted withdioxane (5 mL) and water (1 mL). Ethyl 2-chloropyrimidine-5-carboxylate(164 mg, 1.1 equiv), [1,1′-Bis(diphenylphosphino) ferrocene]dichloropalladium (II) complex with dichloromethane (18 mg, 0.03 equiv),and potassium carbonate (166 mg, 1.5 equiv) were added to the reactionmixture and the resulting suspension was stirred at 80° C. for 2 hoursthen allowed to stand at room temperature for 4 days. The reaction waspartitioned between ethyl acetate (30 mL) and water (50 mL) andextracted with ethyl acetate (2×30 mL). The organic layer was separated,washed with brine (50 mL), dried over sodium sulfate, filtered, andconcentrated to provide crude material that was purified by prep-TLC(5:1 petroleum ether:ethyl acetate) to afford ethyl2-(5-((3-ethoxy-5-fluoropyridin-2-yl)oxy)pyridin-3-yl)pyrimidine-5-carboxylate(50 mg, 16%). MS (ES+) 385.0 (M+H).

Step 4:2-(5-((3-Ethoxy-5-fluoropyridin-2-yl)oxy)pyridin-3-yl)pyrimidine-5-carboxylicacid

Sodium hydroxide (0.20 mL, 3.0 equiv, 2M) was added to ethyl2-(5-((3-ethoxy-5-fluoropyridin-2-yl)oxy)pyridin-3-yl)pyrimidine-5-carboxylate(50 mg, 1.0 equiv) in ethanol (10 mL). The reaction was stirred at 25°C. for 16 hours. The solution was diluted with water (50 mL) andextracted with ethyl acetate (3×30 mL). The aqueous layer was acidifiedwith hydrochloric acid (2N) to a pH of 3. The solution was extractedwith ethyl acetate (15 mL), dried over sodium sulfate, and concentratedto yield2-(5-((3-ethoxy-5-fluoropyridin-2-yl)oxy)pyridin-3-yl)pyrimidine-5-carboxylicacid (25 mg, 54%) as a yellow solid. MS (ES+) 357.0 (M+H).

Step 5:(S)-2-(5-((3-Ethoxy-5-fluoropyridin-2-yl)oxy)pyridin-3-yl)-N-(tetrahydrofuran-3-yl)pyrimidine-5-carboxamide

The title compound was prepared according to general procedure A using2-(5-((3-ethoxy-5-fluoropyridin-2-yl)oxy)pyridin-3-yl)pyrimidine-5-carboxylicacid (25 mg, 1.0 equiv) and (S)-tetrahydrofuran-3-amine (18.3 mg, 3.0equiv). The crude material was purified by prep-HPLC to afford(S)-2-(5-((3-ethoxy-5-fluoropyridin-2-yl)oxy)pyridin-3-yl)-N-(tetrahydrofuran-3-yl)pyrimidine-5-carboxamide(20 mg, 67%) as a white solid. ¹H NMR (400 MHz, DMSO-d6) δ 1.38 (t, 3H),1.87-2.00 (m, 1H), 2.14-2.28 (m, 1H), 3.65 (dd, 1H), 3.70-3.79 (m, 1H),3.83-3.94 (m, 2H), 4.21 (q, 2H), 4.46-4.56 (m, 1H), 7.65-7.74 (m, 2H),8.37 (dd, 1H), 8.65 (d, 1H), 8.98 (d, 1H), 9.29 (s, 2H), 9.40 (d, 1H).MS (ES+) 425.9 (M+H).

Pharmacological Data

The following protocols may of course be varied by those skilled in theart.

Generation of Human DGAT2 (hDGAT2) Construct

A construct for hDGAT2 was generated with an N-terminal FLAG tag (anoctapeptide with the amino acid sequence of AspTyrLysAspAspAspAspLys).For the FLAG-tagged hDGAT2 construct, the cDNA for hDGAT2 wascustom-synthesized at Genscript and cloned into the pFastBac1 vector(Invitrogen) by using BamHI/XhoI restriction enzymes to generate anN-terminally FLAG-tagged pFastBac1-FLAG-hDGAT2 construct (amino acids1-388). The construct was confirmed by sequencing in both directions.

DGAT2 Expression and Preparation of the DGAT2 Membrane Fraction

Recombinant baculovirus for the FLAG-tagged hDGAT2 was generated in SF9insect cells using Bac-to-Bac baculovirus expression system (Invitrogen)according to the manufacturer's protocol. For the expression of hDGAT2,SF9 cells (20 L) grown in Sf900II media were infected with hDGAT2baculovirus at a multiplicity of infection of 1 in a Wave BioreactorSystem 20/50P wave bag (GE Healthcare). After 40 hours of infection, thecells were then harvested by centrifugation at 5,000×g. The cell pelletswere washed by resuspending in phosphate buffered saline (PBS) andcollected by centrifugation at 5,000× g. The cell paste was flash frozenin liquid N₂ and stored at −80° C. until needed. All operations belowwere at 4° C. unless otherwise noted. The cells were resuspended inlysis buffer (50 mM Tris-HCl, pH 8.0, 250 mM sucrose) including 1 mMethylenediaminetetraacetic acid (EDTA) and the complete proteaseinhibitor cocktail (Roche Diagnostics) at a ratio of 3 ml buffer per 1 gcell paste. The cells were lysed by dounce homogenizer. The cell debriswas removed by centrifugation at 1,000×g for 20 min, and the supernatantwas centrifuged at 100,000×g for 1 hour. The resulting pellet was rinsedthree times by filling ultracentrifuge tubes to the top with ice coldPBS before decanting. The washed pellet was resuspended with gentlestirring for 1 hour in lysis buffer containing 8 mM3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) at aratio of 1 mL buffer per 1 g of original cell paste and centrifugedagain at 100,000×g for 1 hour. The resulting supernatant was aliquotted,flash frozen in liquid N₂, and stored at −80° C. until use.

In Vitro DGAT2 Assay and Determination of IC₅₀ Values for DGAT2Inhibitors

For determination of IC₅₀ values, the reactions were carried out in384-well white Polyplates (Perkin Elmer) in a total volume of 20 μL. To1 μL of compounds dissolved in 100% DMSO and spotted at the bottom ofeach well, 5 μL of 0.04% bovine serum albumin (BSA) (fatty acid free,Sigma Aldrich) was added and the mixture was incubated at roomtemperature for 15 minutes. hDGAT2 membrane fractions were diluted in100 mM Hepes-NaOH, pH 7.4, 20 mM MgCl₂ containing 200 nM methylarachidonyl fluorophosphonate (Cayman Chemical; dried from ethyl acetatestock solution under argon gas and dissolved in DMSO as 5 mM stock). 10μL of this enzyme working solution was added to the plates andincubation continued for 2 hours at room temperature. DGAT2 reactionswere initiated by the addition of 4 μL of substrates containing 30 μM[1-¹⁴C]decanoyl-CoA (custom-synthesized by Perkin Elmer, 50 mCi/mmol)and 125 μM 1,2-didecanoyl-sn-glycerol (Avanti Polar Lipids) dissolved in12.5% acetone. The reaction mixtures were incubated at room temperaturefor 40 min and the reactions were stopped by addition of 5 μL of 1%H₃PO₄. After the addition of 45 μL MicroScint-E (Perkin-Elmer), plateswere sealed with Top Seal-A covers (Perkin-Elmer) and phase partitioningof substrates and products was achieved using a HT-91100 microplateorbital shaker (Big Bear Automation, Santa Clara, Calif.). Plates werecentrifuged at 2,000×g for 1 minute in an Allegra 6R Centrifuge (BeckmanCoulter) and then were sealed again with fresh covers before reading ina 1450 Microbeta Wallac Trilux Scintillation Counter (Perkin Elmer).DGAT2 activity was measured by quantifying the generated product[¹⁴C]tridecanoylglycerol in the upper organic phase.

Background activity obtained using 50 μM of(R)-1-(2-((S)-1-(4-Chloro-1H-pyrazol-1-yl)ethyl)-3H-imidazo[4,5-b]pyridin-5-yl)piperidin-3-yl)(pyrrolidin-1-yl)methanone(WO 2013150416, Example 196-A) for complete inhibition of DGAT2 wassubtracted from all reactions. Inhibitors were tested at elevendifferent concentrations to generate IC₅₀ values for each compound. Theeleven inhibitor concentrations employed typically included 50, 15.8, 5,1.58, 0.50, 0.16, 0.05, 0.016, 0.005, 0.0016, and 0.0005 μM. The datawere plotted as percentage of inhibition versus inhibitor concentrationand fit to the equation, y=100/[1+(x/IC₅₀)^(z)], where IC₅₀ is theinhibitor concentration at 50% inhibition and z is the Hill slope (theslope of the curve at its inflection point).

Table 4 below provides the IC₅₀ values of the Examples for inhibition ofDGAT2 in accordance with the above-described assay. Results are reportedas geometric mean IC₅₀ values.

TABLE 4 IC₅₀ values of Examples for inhibition of DGAT2 Example NumberDGAT2 IC₅₀ [nM] 1 17.2 2 200 3.1 13.5 3.2 83.6 3.3 196 3.4 181 3.5 2383.6 35.4 3.7 66.5 4 7.4 5 14.0 6a 20.9 6b 23.2 7 83.0 8 3.7

Determination of IC₅₀ Values for DGAT2 Inhibitors in Human Hepatocytes

For evaluation of the effects of DGAT2 inhibitors in a cell-basedsetting, cryopreserved human hepatocytes (Lot NON and EBS, Celsis,Chicago, Ill.) were thawed and plated onto type I collagen-coated platesaccording to the manufacturer's instructions. After 24 hours overnightrecovery period, the cells were overlayed with media containing 250μg/ml Matrigel (BD Biosciences, San Jose, Calif.). The following day,media was aspirated and replaced with serum-free Williams Media E (LifeTechnologies, Grand Island, N.Y.) containing 400 μM sodium dodecanoate(Sigma-Aldrich, St. Louis, Mo.). Forty-five minutes later, a selectiveDGAT1 inhibitor (Example 3, WO2009016462, prepared as a 100× stocks in25% DMSO, 75% Williams' Media E) was added to all wells at a finalconcentration (3 μM) that completely suppressed endogenous DGAT1activity. DGAT2 inhibitors were then added to the desired finalconcentration. After a 15 minute preincubation, 0.2 μCi[1,3-¹⁴C]-glycerol (American Radio Chemicals, St. Louis, Mo.) was addedto each well followed by a 3 hour incubation. At this point the mediawas removed, the cells washed once with PBS and then lysed in isopropylalcohol: tetrahydrofuran (9:1) prior to centrifugation at 3000 rpm for 5minutes. Radiolabeled lipids were resolved using a 2-solvent system bythin layer chromatography with solvent 1 consisting of ethyl acetate:isopropyl alcohol: chloroform: methanol: 0.25% potassium chloride inwater (100:100:100:40.2:36.1, v/v/v/v) and solvent 2 consisting ofhexane: diethyl ether: acetic acid (70:27:3, v/v/v)). TLC plates weredeveloped in solvent 1 one-third of the plate height, the plate driedunder nitrogen and then developed to the plate top. After separation,radiolabeled lipids were visualized using a Molecular Dynamics'PhosphorImager system. The half maximal inhibitory concentrations (IC₅₀values) were determined using GraphPad Prism (GraphPad Software, Inc.,La Jolla, Calif.) using Hill function with fixed baseline=0 (vehiclecontrol) and Hill slope=1.

In this Setting, Example 1 Showed the Geometric Mean IC₅₀ Value of 2.8nM (N=10)

In Vivo effects of DGAT2 inhibitors on plasma and hepatic triglyceridelevels

The rat western diet model was utilized to assess the longer termeffects of the treatment with DGAT2 inhibitors on plasma triglycerideproduction and hepatic triglyceride content. Male Sprague-Dawley ratswere housed under standard laboratory conditions on a 12-hour light,12-hour dark cycle (lights on at 06:00). Two weeks prior to study startanimals were placed on a high-fat, high-cholesterol diet (D12079b,Research Diets, New Brunswick, N.J.). This diet provides ˜43% ofkilocalories from carbohydrate and ˜41% of kilocalories from fat. DGAT2inhibitors were administered orally as a solution (10 mL/kg dosingvolume) in 0.5% HPMCAS-HF and 0.015% SLS in DI water, pH 8.5(methylcellulose and butylated hydroxytoluene were obtained fromSigma-Aldrich, St. Louis, Mo.). Vehicle-treated animals received anaqueous solution of 0.5% HPMCAS-HF and 0.015% SLS in DI water, pH 8.5alone. DGAT2 inhibitors were administered orally twice daily for 7 daysat 08:00 and 16:00 at 1, 3, 10, 30 and 90 mg/kg. On day 8, all animalswere fasted at 06:00, dosed with vehicle or DGAT2 inhibitors at 10:00and sacrificed 2 hours post-dose. Rats were sacrificed by carbon dioxideasphyxiation and blood collected via lateral tail vein. Plasma TG levelswere determined using a Roche Hitachi Chemistry analyzer according tothe manufacturer's instructions (Roche Diagnostics Corporation,Indianapolis, Ind.) and data was analyzed using GraphPad Prism (GraphPadSoftware, Inc., La Jolla, Calif.). Livers sample collection fordetermination of hepatic triglyceride content was excised at time ofsacrifice, immediately frozen in liquid nitrogen, and held at −80° C.until analysis. For assessment of tissue triglyceride levels a sectionof liver wrapped in aluminum foil was pulverized with a hammer, on analuminum heat block in a liquid nitrogen bath. Pulverization of theliver tissue produced a homogeneous powder. Homogenization buffer, TrispH 7.4, 98.9 milliliters 0.9% NaCl and 100 microliters of Triton X 100,was mixed on a stir plate for 10 minutes prior to using. Sample weightsof approximately one-hundred milligrams of homogenous liver tissue wereweighed and placed in Lysing Matrix D tube (MP Biomedicals, Cat#6913-100) with 1 mL of homogenization buffer. All samples were thenplaced in the FastPrep FP120 (MP Biomedicals, Cat #6001-120) for 2minutes or until tissue was properly homogenized. All samples were thenspun for 30 seconds at 10,000 g, to clear foam from homogenization. 50microliters of sample was transferred to a sterile mixing plate with 450microliters of Dulbeccos phosphate-buffered saline (DPBS) to create a1:10 dilution. Upon re-suspension of the new sample, all samples weretransferred to sampling tubes for the Siemens Advia XPT ClinicalAnalyzer. The triglyceride assay was performed through absorbance andreported as milligrams per deciliter. Triglycerides were then normalizedper gram of tissue in Microsoft Excel.

FIGS. 3 and 4 summarize the effects of oral administration with Example1 on plasma and hepatic triglyceride levels in western diet fed SpragueDawley rats in accordance with the above-described methods. Data aremean±standard deviation from 8 animals. Difference between group meansrelative to vehicle was performed by a 1-way ANOVA followed by aDunnett's multiple comparisons test **p<0.01, ****p<0.0001.

Throughout this application, various publications are referenced. Thedisclosures of these publications in their entireties are herebyincorporated by reference into this application for all purposes.

It will be apparent to those skilled in the art that variousmodifications and variations can be made in the present inventionwithout departing from the scope or spirit of the invention. Otherembodiments of the invention will be apparent to those skilled in theart from consideration of the specification and practice of theinvention disclosed herein. It is intended that the specification andexamples be considered as exemplary only, with a true scope and spiritof the invention being indicated by the following claims.

What is claimed is:
 1. A method for the reduction of at least one pointin severity of nonalcoholic fatty liver disease or nonalcoholicsteatohepatitis grading scoring systems, reduction of the level of serummarkers of nonalcoholic steatohepatitis activity, reduction ofnonalcoholic steatohepatitis disease activity or reduction in themedical consequences of nonalcoholic steatohepatitis in humanscomprising the step of administering to a human in need of suchreduction an effective amount of a compound according to Formula (I) ora pharmaceutically acceptable salt of said compound to a patient in needthereof; wherein the compound of Formula (I) is:

wherein D¹ and D² are each independently N or CH; R¹ is H, or(C₁-C₂)alkyl optionally substituted with one or two substituents eachindependently selected from fluoro and (C₃-C₆)cycloalkyl; R² is H orfluoro; R³ is,

R⁴ is H, cyano, or (C₁-C₄)alkyl optionally substituted with one or twosubstituents each independently selected from —OH and —S(O)₂R⁶; R⁵ is Hor —OH; and R⁶ is (C₁-C₄)alkyl.
 2. A method for treating hyperlipidemia,Type I diabetes, Type II diabetes mellitus, idiopathic Type I diabetes(Type Ib), latent autoimmune diabetes in adults (LADA), early-onset Type2 diabetes (EOD), youth-onset atypical diabetes (YOAD), maturity onsetdiabetes of the young (MODY), malnutrition-related diabetes, gestationaldiabetes, coronary heart disease, ischemic stroke, restenosis afterangioplasty, peripheral vascular disease, intermittent claudication,myocardial infarction, dyslipidemia, post-prandial lipemia, conditionsof impaired glucose tolerance (IGT), conditions of impaired fastingplasma glucose, metabolic acidosis, ketosis, arthritis, obesity,osteoporosis, hypertension, congestive heart failure, left ventricularhypertrophy, peripheral arterial disease, diabetic retinopathy, maculardegeneration, cataract, diabetic nephropathy, glomerulosclerosis,chronic renal failure, diabetic neuropathy, metabolic syndrome, syndromeX, premenstrual syndrome, angina pectoris, thrombosis, atherosclerosis,transient ischemic attacks, stroke, vascular restenosis, hyperglycemia,hyperinsulinemia, hypertrygliceridemia, insulin resistance, impairedglucose metabolism, erectile dysfunction, skin and connective tissuedisorders, foot ulcerations and ulcerative colitis, endothelialdysfunction and impaired vascular compliance, hyper apo Blipoproteinemia, Alzheimer's, schizophrenia, impaired cognition,inflammatory bowel disease, ulcerative colitis, Crohn's disease, andirritable bowel syndrome, non-alcoholic steatohepatitis (NASH), ornon-alcoholic fatty liver disease (NAFLD), in humans comprising the stepof administering to a human in need of such treatment a therapeuticallyeffective amount of a compound according to Formula (I) or apharmaceutically acceptable salt of said compound; wherein the compoundof Formula (I) is:

wherein D¹ and D² are each independently N or CH; R¹ is H, or(C₁-C₂)alkyl optionally substituted with one or two substituents eachindependently selected from fluoro and (C₃-C₆)cycloalkyl; R² is H orfluoro; R³ is,

R⁴ is H, cyano, or (C₁-C₄)alkyl optionally substituted with one or twosubstituents each independently selected from —OH and —S(O)₂R⁶; R⁵ is Hor —OH; and R⁶ is (C₁-C₄)alkyl.
 3. The method of claim 1 wherein themethod reduces portal hypertension, hepatic protein syntheticcapability, hyperbilirubinemia, or encephalopathy.